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Bioenergetic therapy based on tumor glucose metabolism is emerging as a promising therapeutic modality. To overcome the poor bioavailability and toxicity of arenobufagin (ArBu), a MOF-derived intelligent nanosystem, ZIAMH, was designed to facilitate energy deprivation by simultaneous interventions of glycolysis, OXPHOS and TCA cycle. Herein, zeolitic imidazolate framework-8 was loaded with ArBu and indocyanine green, encapsulated within metal–phenolic networks for chemodynamic therapy and hyaluronic acid modification for tumor targeting. ZIAMH nanoparticles can release ArBu in the tumor microenvironment for chemtherapy, and ICG enables photothermal therapy under near-infrared laser irradiation. In vitro and in vivo mechanism studies revealed that the ZIAMH nanoplatform downregulated glucose metabolism related genes, resulting in the reduction of energy substances and metabolites in tumors. Additionally, it significantly promoted cell apoptosis by upregulating pro-apoptotic proteins such as Bax, Bax/Bcl-2, cytochrome C. Animal studies have shown that the tumor inhibition efficiency of ZIAMH nanomedicines was three fold higher than that of free drugs. Therefore, this study provides a new strategy for glucose metabolism-mediated bioenergetic therapy and PTT/CDT/CT combined therapy for tumors. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate the direct effects of α-Linolenic acid (ALA) on the in vitro production of testosterone and the expression of key enzymes and proteins related to steroidogenesis in Leydig cells of roosters. Purified primary Leydig cells isolated from 65-week-old roosters were purified and treated with different concentrations of ALA treatments: (0 μm/L [control], solvent control group (DMSO), 20 μM/L, 40 μM/L, and 80 μM/L) and cell counting-8 (CCK-8) for cell viability assay, Enzyme-linked immunosorbent assay (ELISA) kit for the determination of testosterone in cell supernatants, quantitative (real-time) PCR, and analysis of activities of antioxidants catalase (CAT), superoxide dismutase (SOD) and malondialdehyde (MDA), evaluation of mitochondrial membrane potential, pro- and anti-apoptotic proteins/genes Bcl-2, Bcl-2-associated X protein (Bax), apoptosis-inducing factor (AIF) were done respectively. Our results showed that ALA significantly increased testosterone secretion in primary rooster Leydig cells (P < 0.05), and 40 μM/L is the optimal dose. Leydig cells supplemented with ALA (20, 40, 80 μM) increased the expression of key enzymes and proteins 3β-hydroxysteroid dehydrogenase (3β-HSD), steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc) concerning steroidogenesis, enhanced antioxidant capability, improved mitochondrial biogenesis, and markedly improved the mitochondrial membrane potential (P < 0.05). Furthermore, the expression of the apoptosis-suppressive gene Bcl-2 was significantly increased, but Bax and AIF expression was decreased in the ALA group compared to that in the control group (P < 0.05). ALA promoted testosterone production, enhanced steroidogenic enzyme expression, improved mitochondrial function, and antioxidant capacity, and reduced apoptosis in primary rooster Leydig cells, with 40 μM/L identified as the optimal concentration. [ABSTRACT FROM AUTHOR]

• FMT improves neurological function, reduces infarct volume in stroke rats. • FMT inhibits necroptosis and apoptosis in ischemic brain tissue. • FMT reduces iNOS-positive microglia activation in ischemic brain. • FMT may be a therapy for stroke by modulating cell death and inflammation. This study aims to delve into the mechanisms underlying the improvement of neurological function in rats with ischemic stroke through fecal microbiota transplantation. A total of fifty male Sprague-Dawley rats were categorized into four groups: Sham, MCAO, MCAO+vehicle and FMT. We assessed behavioral and pathological alterations in the rats using modified neurological function scoring and TTC staining.Additionally, Western blot and immunofluorescence were used to detect the expression levels of Apoptotic and Necroptosis markers in neurons of ischemic brain tissue, and immunofluorescence was used to analyze the degree of activation of microglia. FMT group exhibited a decline in neurological function score compared to the MCAO and MCAO + vehicle group, accompanied by a reduction in infarct volume (P < 0.05). Relative to the SHAM group, the MCAO group displayed a significant increase in the expression levels of necroptosis-related proteins Phospho-RIP1, Phospho-RIP3, Phospho-MLKL, apoptotic proteins Bax and Cleaved caspase-3, and the iNOS positive microglia in ischemic brain tissue, while Bcl-2 expression was notably decreased (P < 0.05).Conversely, compared to the MCAO + vehicle group, the FMT group showed decreased expression levels of Phospho-RIP1, Phospho-RIP3, Phospho-MLKL, Bax, Cleaved caspase-3, and iNOS-positive microglia, while the expression of Bcl-2 was increased. Fecal microbiota transplantation offers a promising approach to improving neurological function in rats with ischemic stroke by inhibiting neuronal apoptosis, necroptosis, and the polarization of inflammatory microglial cells. [ABSTRACT FROM AUTHOR]

The article "Correction to 'Role and Regulation of Proapoptotic Bax in Oral Squamous Cell Carcinoma and Drug Resistance'" discusses errors in the images of Bax protein expression and p‐Akt expression in oral squamous cell carcinoma. The correction provides the accurate versions of the figures without changing the conclusions of the original article. The study examines the role of Bax in oral squamous cell carcinoma progression and resistance to chemoradiation therapy, highlighting the correlation between Bax protein expression, Bax mRNA expression, and Bax promoter methylation. [Extracted from the article]

Background: Drug resistance remains a significant obstacle to Acute myeloid leukemia (AML) successful treatment, often leading to therapeutic failure. Our previous studies demonstrated that Glioma-associated oncogene-1 (GLI1) reduces chemotherapy sensitivity and promotes cell proliferation in AML cells. GANT61, an inhibitor of GLI1, emerges as a promising candidate in AML treatment. This study aims to explore the effects of the combination of GANT61 and Adriamycin (ADR) on AML cells resistance and elucidate the mechanisms through which GANT61 may potentiate the sensitivity of AML cells to ADR. Methods: AML cell lines and AML primary cells were studied to evaluate effects and mechanisms of GANT61. Flow cytometry assays were used to verify apoptosis. Cell Counting Kit-8 (CCK-8) and EDU+ staining were used to observe changes in cell viability and the cytotoxic effect to different drugs. The transcriptomic profiles of HL-60/ADR cells with or without GANT61 treatment were compared via RNA-Seq analysis. Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses and Gene Set Enrichment Analysis (GSEA) were performed for differentially expressed genes (DEGs) to reveal the underlying mechanisms of GANT61 in AML cells. GLI1, BCL2, Bax protein and mRNA expression levels were assessed by Western blot and Real-time polymerase chain reaction (RT-PCR). Results: Our studies found that the combination of GANT61 and ADR synergistically inhibits proliferation while enhancing apoptosis in HL-60/ADR cells, and does not significantly exacerbate myelosuppression. Mechanistically, GSEA revealed enrichment of the gene set associated with the KEGG term "Apoptosis" and "Lysosome" in GANT61 treated cells. Meanwhile, "Apoptosis" was identified as the third most relevant pathway enriched by lysosomal DEGs, and BCL2 expression showed a negative correlation with these lysosomal DEGs in AML patients. RT-PCR and Western blot analysis disclosed that GANT61 significantly restrained BCL2 expression in AML cells. Lastly, we proved that venetoclax, a BCL2 inhibitor, co-treatment with GANT61 improved ADR sensitivity in HL-60/ADR cells. Conclusions: GANT61 effectively reversed ADR resistance in HL-60/ADR cells by upregulating lysosome activities and downgrading BCL2 expression, providing a new treatment strategy with acceptable toxicity for AML-resistant patients. [ABSTRACT FROM AUTHOR]

Background: Cardiac contractility modulation (CCM) is non-excitatory electrical stimulation for improving cardiac function. This study aimed to evaluate the effects of CCM on autophagy and apoptosis of cardiac myocytes in a rabbit model of chronic heart failure (CHF) and explore its possible mechanism. Methods: Thirty rabbits were randomised into the Sham, heart failure (HF) and CCM groups, and animals in all three groups were sacrificed after 16 weeks of ascending aortic constriction or sham surgery. The expression of autophagy associated protein LC3 was observed by immunofluorescence staining. With Western-blot measured the expression of Beclin1, P62, LC3B (II/I) and Bcl-2, ALDH2, Bax and Caspase-3 protein in myocardial tissue. The apoptosis rate and the apoptosis of myocardial cells was observed by flow cytometry and TUNEL method. Results: 1) In comparison to the Sham group, the expression of LC3 and Beclin1 was significantly increased, and the expression of p62 protein was decreased in the heart tissues of rabbits in the HF group. Compared with HF group, after CCM intervention, the expression of Beclin1 and LC3B proteins decreased, while the P62 protein increased, and the LC3B(II/I) ratio decreased (P<0.05). 2) The expression of Bcl-2, ALDH2 protein and Bcl-2 mRNA decreased compared with the Sham group (P<0.05), while the expression of Bax, Caspase-3 protein and mRNA was significantly increased (P<0.05). However, the expression of ALDH2 mRNA in the CCM group was not statistically significant. The expression of Bcl-2, ALDH2 protein and mRNA increased after CCM intervention, and the expression of Bax, Caspase-3 protein and mRNA decreased (P<0.05). 3) The apoptosis situation in the Sham group was similar to that of normal myocardium, compared with the Sham group, the number of apoptotic bodies increased, and the apoptosis percentage of cardiomyocytes increased significantly (P<0.05). After CCM intervention, the number of apoptotic bodies and the percentage of apoptosis decreased compared with the HF group (P<0.05). Conclusions: The intervention of CCM has been shown to enhance both myocardial systolic and diastolic function in rabbits with CHF. The mechanism may be related to the inhibition of cardiomyocyte autophagy by regulating the expression levels of Beclin1, P62, and LC3B(II/I) in cardiomyocytes, as well as the reversal of cardiomyocyte apoptosis by regulating the expression levels of Bcl-2, ALDH2, Bax, and Caspase-3 in cardiomyocytes. [ABSTRACT FROM AUTHOR]

Cumin, a popular spice, is widely used to treat stomach ailments in Central Asia and Xinjiang, China. Cumin essential oil has been found to effectively treat gastric ulcers, but its pharmacodynamic basis remains unclear. In this study, cumin essential oil was directly separated using column chromatography, and its components were identified through multi‐dimensional gas chromatography‐mass spectrometry. Finally, the cumin essential oil was fractionated into E1, E2, and E3. The effects of these fractions on gastric ulcers were studied using an anhydrous ethanol‐induced rat model. The results indicated that the three fractions decreased ulcer index, gastric fluid pH, and pepsin activity to different extents. They lowered the levels of prostaglandin E2, gastrin, and epidermal growth factor in rat serum. According to an analysis of the above indices, E3 fraction had the best anti‐ulcer effect. The detection results of the oxidative stress and inflammatory factors showed that all three fractions relieved the ethanol‐induced oxidative stress and reduced the release of inflammatory factors to varying degrees. The E3 fraction played the most significant role. The E3 fraction was selected to explore the relevant mechanism, and the results showed that E3 fraction significantly prevented the cleaved caspase‐3 and Bax protein levels that were ethanol‐induced and resisted apoptosis induced by ethanol injury. The western blot results for detecting the NF‐κB‐related pathway protein showed that E3 fraction significantly inhibited the activation of p‐p65, p‐IKKβ, and p‐IκBα. The study found that the E3 fraction of cumin essential oil had the most effective anti‐ulcer effect by inhibiting NF‐κB activation and apoptosis, thus reducing inflammation. [ABSTRACT FROM AUTHOR]

Objective: This study examines the therapeutic potential of monotropein (Mon) in a rat model of acute pulmonary embolism (APE), aiming to elucidate its mechanistic role and provide new insights for APE treatment. Methods: Thirty Sprague Dawley (SD) rats were randomly assigned to five groups (n = 6 per group): sham, Mon (40 mg/kg), APE, APE + 20 mg/kg Mon, and APE + 40 mg/kg Mon. APE was induced via autologous thrombus infusion in all groups except sham and Mon-only groups. We assessed blood gas parameters, lung wet/dry weight (W/D) ratio, and oxidative stress markers. Additionally, excised lung tissues underwent evaluation for serum inflammatory factors via ELISA, apoptotic cells via TUNEL assay, and protein expression via Western blot. Results: Compared to the sham group, APE-induced rats exhibited significantly elevated blood oxygen levels and increased pro-inflammatory factors, including interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and IL-8. Mon treatment effectively mitigated these APE-induced changes, reducing blood oxygen concentration and downregulating IL-1β and TNF-α levels. Furthermore, Mon demonstrated anti-apoptotic effects by decreasing cleaved caspase-3 and Bax protein levels while upregulating Bcl-2 expression. Mon also suppressed nuclear factor-κB (NF-κB) activation by inhibiting the phosphorylation levels of p65/RelA and IκBα proteins, while the total protein level of IκBα was increased with Mon treatment. Conclusion: Mon effectively ameliorated lung tissue injury in APE rats by inhibiting apoptosis, attenuating inflammatory responses, and alleviating oxidative stress. These beneficial effects appear to be mediated through modulation of the NF-κB pathway, suggesting Mon as a promising therapeutic candidate for APE treatment. [ABSTRACT FROM AUTHOR]

Objective: To investigate the mechanism of overexpression of miR-320e in inhibiting inflammatory response of respiratory syncytial virus (RSV) infected bronchial epithelial cells. Methods: Human bronchial epithelial cells 16HBE were cultured in vitro and infected with RSV, and cells were divided into Con group, RSV group, RSV+miR-NC group, RSV+miR-320e group, RSV+miR-320e+vector group, RSV+miR-320e+TLR4 group. RT-qPCR was used to detect expression levels of miR-320e and TLR4 mRNA; MTT to detect cell proliferation changes; flow cytometry to detect cell apoptosis; Western blot was used to detect Bcl-2, Bax, TLR4, IκBα, p-IκBα, NF-κB and p-NF-κB protein expressions; ELISA to detect TNF-α, IL-6, IL-1β and IFN-α, IFN-β expressions; dual luciferase experiment to verify the tageting relationship between miR-320e and TLR4. Results: Compared with Con group, miR-320e expression level, survival rate, Bcl-2 and IκBα protein expressions were significantly reduced, apoptosis rate, Bax protein expression, TNF-α, IL-6, IL-1β and IFN-α, IFN-β expressions, TLR4 mRNA and protein expression, and p-IκBα protein expression and p-NF-κB/NF-κB were increased significantly in RSV group. Compared with RSV+miR-NC group, miR-320e expression level, survival rate, IFN-α, IFN-β expressions, Bcl-2 and IκBα protein expressions were significantly increased, apoptosis rate, Bax protein expression, TNF-α, IL-6, IL-1β expressions, TLR4 mRNA and protein expression, and p-IκBα protein expression and p-NF-κB/NF-κB in RSV+miR-320e group were significantly reduced. miR-320e targets and negatively regulates the expression of TLR4. Upregulation of TLR4 can partially restore the effect of overexpression of miR-320e on apoptosis and inflammatory response of RSV-infected bronchial epithelial cells 16HBE. Conclusion: miR-320e inhibits 16HBE apoptosis and inflammation in RSV-infected bronchial epithelial cells by targeting and negatively regulating TLR4 expression. [ABSTRACT FROM AUTHOR]

Objective: To study whether and, if so, how honokiol overcome dexamethasone resistance in DEX-resistant CEM-C1 cells. Methods: We investigated the effect of honokiol (0–20 µM) on cell proliferation, cell cycle, cell apoptosis and autophagy in DEX-resistant CEM-C1 cells and DEX-sensitive CEM-C7 cells. We also determined the role of c-Myc protein and mRNA in the occurrence of T-ALL associated dexamethasone resistance western blot and reverse transcription-qPCR (RT-qPCR) analysis. Results: Cell Counting Kit (CCK)−8 assay shows that DEX-resistant CEM-C1 cell lines were highly resistant to dexamethasone with IC50 of 364.1 ± 29.5 µM for 48 h treatment. However, upon treatment with dexamethasone in combination with 1.5 µM of honokiol for 48 h, the IC50 of CEM-C1 cells significantly decreased to 126.2 ± 12.3 µM, and the reversal fold was 2.88. Conversely, the IC50 of CEM-C7 cells was not changed combination of dexamethasone and honokiol as compared to that of CEM-C7 cells treated with dexamethasone alone. It has been shown that honokiol induced T-ALL cell growth inhibition by apoptosis and autophagy via downregulating cell cycle-regulated proteins (Cyclin E, CDK4, and Cyclin D1) and anti-apoptotic proteins BCL-2 and upregulating pro-apoptotic proteins Bax and led to PARP cleavage. Honokiol may overcome dexamethasone resistance in DEX-resistant CEM-C1 cell lines via the suppression of c-Myc mRNA expression. Conclusion: The combination of honokiol and DEX were better than DEX alone in DEX-resistant CEM-C1 cell lines. Honokiol may regulate T-ALL-related dexamethasone resistance by affecting c-Myc. [ABSTRACT FROM AUTHOR]

Aristolochic acid I (AAI) is strongly nephrotoxic and can cause "Aristolochic acid nephropathy (AAN)". Aristolochic acid nephropathy is characterized by extensive renal interstitial fibrosis. However, the exact mechanism by which it occurs has not been fully elucidated. lt has been reported that indoleamine 2,3-dioxygenase-1 (IDO1) promotes renal fibrosis in renal disorders, but it is unclear how IDO1 functions in AAI-induced kidney fibrosis. In this work, we systematically examined the role of IDO1 in AAI-induced renal tubulointerstitial fibrosis. The results showed that AAI induced upregulation of IDO1 expression in renal tubular epithelial cells and mouse kidney. Inhibition of IDO1 expression reduced the levels of fibrosis-associated markers α-SMA, COL-I and FN and ameliorated renal tubular epithelial cell fibrosis. It also improved renal function, reduced collagen deposition, and ameliorated interstitial fibrosis in mice. Moreover, we discovered that inhibition of IDO1 decreased the expression of the apoptotic protein BAX, raised the expression of BCL-2 protein, and reduced apoptosis. The above studies suggest that IDO1 is a target of action in renal tubulointerstitial fibrosis caused by AAI, and inhibition of IDO1 may be a viable approach for the therapy of AAI-induced renal tubulointerstitial fibrosis. [Display omitted] • The role of IDO1 in AAI-induced renal interstitial fibrosis were first verified. • The present study provides a new target for the therapy of AAI-induced renal tubulointerstitial fibrosis as well as a new direction into the mechanism of AAI-induced renal tubulointerstitial fibrosis. [ABSTRACT FROM AUTHOR]

Context: Cisplastin (CDDP) is a chemotherapeutic drug frequently used to manage a variety of cancers. However, its use is associated with hepatorenal toxicity resulting from elevated reactive oxygen species production. Objective: Herein, the hepatorenal protective effect of tert-butylhydroquinone (tBHQ) in cisplatin (CDDP)-treated rats was examined. Methods: Wistar male rats randomly divided into four groups: normal control, tBHQ, CDDP and tBHQ + CDDP received 50 mg/kg b.w./day of tBHQ orally for 14 days while 7 mg/kg b.w of CDDP was administered intraperitoneally on Day 8. Results: CDDP increased serum biomarkers of hepatic (AST, ALP, ALT, GGT) and renal (creatinine, urea, uric acid, kidney injury molecule 1) function. The levels of nuclear factor erythroid-2-related factor 2 protein and the activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase activities were decreased in liver and kidney. Also, CDDP increased hepatic and renal levels of NF-κB, TNFα, Bax and caspase-3 proteins and decreased hepatorenal levels of Bcl-2 protein in the liver and kidney. Pre-treatment with tBHQ prevented these negative effects. Significance: Pre-intervention with tBHQ attenuates hepatorenal toxicity of CDDP by dampening oxidative stress, inflammation and apoptosis. [ABSTRACT FROM AUTHOR]

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Molecular analysis of breast cancer cells by niosomal encapsulated Withania somnifera (WS) extract is the focus of this investigation. Characterization of ASH-loaded niosomes produced by a thin film technique showed that they have an appropriate zeta potential, small particle size, and low polydispersity index. The concentration-dependent suppression of cell proliferation was seen in MCF-7 breast cancer cells treated with ASH-NIO, with an IC50 of 24.13 µM, and olaparib at values of 10.45 µM. Molecular investigation showed that ASH-NIO treated cells had lower levels of B-cell lymphoma 2 (Bcl2) mRNA expression compared to untreated cells, and upregulated levels of Tumor protein 53 (P53), Bcl-2-associated X protein (Bax), caspase-9, and caspase-3. These results provide more evidence that ASH-NIO has potential anti-cancer effects and may be useful as an adjunctive treatment for breast cancer. [ABSTRACT FROM AUTHOR]

Glioma is a common and fatal malignant primary brain tumor. Radiotherapy and first-line chemotherapy have little effect on the survival rate of patients, requiring alternative therapies. The main active ingredient of Tripterygium wilfordii Hook. F. triptolide (TP) has been shown to have anti-inflammatory and anti-proliferative properties, along with a wide range of anticancer activities. This study aimed to investigate the molecular mechanisms of triptolide in glioma treatment through network pharmacology and experimental validation. Cell viability was first assessed using Cell Counting Kit-8 (CCK8), followed by cell scratch assay and cell migration ability. Apoptosis-related markers, including TUNEL staining, Bcl-2-associated X protein (Bax), and B-cell lymphoma-2 (Bcl-2), were detected. Network pharmacology was used to predict the key targets of glioma, detect its signal pathways, screen the key components and targets for molecular docking, and explore the signaling pathways of TP. Lastly, immunofluorescence assays and ELISA were performed to elucidate the underlying mechanistic pathways. The network pharmacology data suggested that TP may inhibit glioma proliferation by regulating the signaling pathway of the nuclear factor kappa-B (NF-κB). The results showed that the underlying mechanism involved the regulation of the NF-κB signaling pathway to promote the generation of reactive oxygen species, thereby enhancing oxidative stress response and promoting cell apoptosis. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of (+)-borneol on neuroinflammation and microglia phenotype polarization in epileptogenesis and its possible mechanism. Methods: Based on mouse models of status epilepticus (SE) induced by pilocarpine, and treated with 15 mg/kg (+)-borneol, western-blot was used to detect the expressions of NeuN, Iba-1, TLR4, p65 and p-p65 in the hippocampus. Immunofluorescence was used to detect the expression of apoptosis-related proteins Bax and Bcl-2. To explore the effect of (+)-borneol on microglia in vitro , we used the kainic acid-induced microglia model and the concentration of (+)-borneol was 25 μM according to CCK-8 results. The levels of tumor necrosis factor- α (TNF-α), interleukin-1β (IL-1β) and interleukin-10 (IL-10) in the supernatant of each group was detected by ELISA. The nitric oxide (NO) content in the supernatant was detected by Griess method. The expressions of Iba-1 and TLR4-NFκB signaling pathway-related proteins (TLR4, p65, p-p65) were detected by Western-Blot. Immunofluorescence was used to detect microglia's M1 and M2 phenotype polarization and the expression of Iba-1 and TLR4. Results: (+)-borneol reduced hippocampal neuronal injury, apoptosis, and microglia activation by inhibiting the TLR-NFκB signaling pathway in SE mice. TLR4 agonist LPS partially reversed the neuroprotective effect of (+)-borneol. In the KA-induced microglia model, (+)-borneol inhibited microglia activation, M1 phenotype polarization, and secretion of pro-inflammatory cytokines through the TLR4-NFκB signaling pathway. LPS treatment inhibited the therapeutic effects of (+)-borneol. Conclusion: (+)-borneol inhibits microglial neuroinflammation and M1 phenotype polarization through TLR4-NFκB signaling pathway and reduces neuronal damage and apoptosis in SE mice. Therefore, (+)-borneol may be a potential drug for epilepsy modification therapy. [ABSTRACT FROM AUTHOR]

2H‐pyran compound 1 synthesized from 6‐methylpyridine‐2,4‐diol and citral, has been used for Cu‐catalyzed N‐arylation with a range of arylboric acids to obtain arylated pyranopyridine core structure derivatives (yield up to 77 %). Among them, compound 3 h exhibited a much better inhibitory effect on HepG2 liver cancer cells compared to citral, and the IC50 value was 5.3 μM following exposure with the newly synthesized derivatives (herein named 3 h for short in this paper), which was lower than that of the cisplatin (6.5 μM). Meanwhile, the cell‐cycle arrest of HepG2 cells occurred in the S phase, and the apoptosis of HepG2 cells was significantly increased with increasing drug concentration. In addition, real‐time fluorescence quantification PCR and Western blotting experiments showed that the expression of apoptotic protein BAX was increased, while the expression of anti‐apoptotic protein BCL2 was inhibited in a dose‐dependent fashion. The results of these experiments indicated that apoptosis was promoted in HepG2 cells via apoptotic signaling pathway activated by 3 h. Furthermore, 3 h effectively decreased the phosphorylation levels of PI3 K, ATK and ERK, resulting in the inhibitions of MAPK/ERK and PI3 K/ATK signaling pathways. Briefly, 3 h has been found to show inhibitory effects on the survival of HepG2 liver cancer cells and may be used as anti‐cancer drug in the future. [ABSTRACT FROM AUTHOR]

Introduction: Interleukin-41 (IL-41), also known as Metrnl, is a multifunctional adipokine recognized for its neurotrophic and anti-inflammatory properties that play a significant role in diseases such as sepsis and chronic obstructive pulmonary disease. Despite its crucial biological functions, the mechanisms by which IL-41 mitigates lipopolysaccharide (LPS)-induced acute lung injury (ALI) are not well understood. This study aimed to elucidate the protective effects of IL-41 against LPS-induced inflammation and apoptosis in a murine model and in vitro using bronchial epithelial cells (Beas-2b). Methods: We administered recombinant IL-41 to mice subjected to LPS-induced ALI and observed changes in lung histopathology, inflammation, and apoptosis. Concurrently, Beas-2b cells were transfected with IL-41 constructs, and the role of the AMPK/SIRT1 pathway was investigated using specific inhibitors and agonists. Results: Our results demonstrated that LPS-induced ALI is characterized by increased inflammatory cell chemotaxis in lung lavage fluid, enhanced phosphorylation of NFκB p65, and elevated Bax protein expression, coupled with a decrease in IL-41 protein levels. Treatment with recombinant IL-41 effectively mitigated these pathological changes by upregulating AMPK phosphorylation and SIRT1 expression and inhibiting IκB/NFκB p65 phosphorylation. In cellular assays, overexpression of IL-41 reversed LPS-induced oxidative stress, apoptosis, and the secretion of inflammatory cytokines, whereas suppression of IL-41 or inhibition of AMPK reversed these protective effects. Conclusions: In conclusion, IL-41 exerts significant protective effects against ALI by activating the AMPK/SIRT1 signaling pathway and reducing excessive inflammation and apoptosis. These findings suggest that IL-41 holds promise as a therapeutic target for ALI, potentially allowing for personalized treatments based on serum IL-41 levels. [ABSTRACT FROM AUTHOR]

Fusarium sacchari is a significant pathogenic fungus that causes sugarcane Pokkah Boeng. Proteins secreted by pathogenic fungi can be delivered into hosts to suppress plant immunity and establish infection. However, there is still much to be discovered regarding F. sacchari's secreted effectors in overcoming plant immunity. In this paper, we characterize a novel effector called FsMEP1, which is essential for the virulence of F. sacchari. FsMEP1 contains a conserved zinc-binding motif sequence, HEXXH, and is highly expressed during host infection. Using the Agrobacterium tumefaciens-mediated transient expression system, it was confirmed that FsMEP1 could suppress Bcl-2-associated X protein (BAX)-triggered cell death, callose deposition, and ROS explosion in Nicotiana benthamiana. Furthermore, the deletion of FsMEP1 demonstrated its requirement for contributing to the pathogenicity of F. sacchari in sugarcane. Further analysis revealed that FsMEP1 could interact with the sugarcane thiamine thiazole synthase ScTHI2 and disrupt its normal localization, thereby inhibiting the synthesis of thiamine and the defense responses mediated by ScTHI2. Based on these findings, we propose that ScTHI2 represents a potential molecular target for improving sugarcane resistance to Pokkah Boeng disease. [ABSTRACT FROM AUTHOR]

In this study, water-soluble polysaccharides purified from burdock root were used to intervene in carbon tetrachloride (CCl4)-induced acute liver injury (ALI) of BRL3A hepatocytes and rats. Our results indicated that CCl4 significantly inhibited hepatocyte viability and upregulated the expression of reactive oxygen species (ROS), malondialdehyde (MDA), pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6), and the pro-apoptotic protein Bax. However, Arctium lappa L. root polysaccharides (ALP) could effectively ameliorate liver function and histopathology, oxidative stress, and inflammatory markers. In addition, ALP reduced the expression of apoptotic markers and promoted the proliferation of damaged hepatocytes. In conclusion, ALP possesses a hepatoprotective effect mediated by attenuating oxidative damage, inflammation and apoptosis in ALI. [ABSTRACT FROM AUTHOR]

Purpose To explore the effect of Huashirunzao prescription on the Hippo-Yes associated protein (YAP)/transcriptional coactivator with PDZ binding motif (TAZ) pathway on submandibular gland cell apoptosis and salivary secretion function in Sjögren's syndrome (SS) model mice (NOD/Ltj). method Eight-week-old female BALB/c mice were selected as the blank group, and eight-week-old NOD/Ltj female mice were randomly divided into the model group, hydroxychloroquine group, and low-, medium-, and high-dose groups of the drug. After 8 weeks of administration, the water intake and salivary flow rate of the mice in each group were measured. The pathological damage of the submandibular gland tissues of the mice in each group was detected by hematoxylin-eosin staining. The expressions of aquaporin 5 (AQP5), B-cell lymphoma/leukemia-2 protein (Bcl-2), Bcl-2-associated X protein (Bax), beclin-1 protein (beclin-1), YAP, phosphorylated YAP (p-YAP), and TAZ proteins in the submandibular gland tissues were detected by immunohistochemistry and Western blotting. The apoptosis rate of submandibular gland cells in each group was detected by TUNEL staining. result Compared with the blank group, the Hippo-YAP/TAZ pathway was inhibited in the submandibular gland tissue of mice in the model group, and the lymphocyte infiltration increased, the water intake increased, the salivary flow rate decreased, the AQP5 protein expression decreased, and the apoptosis rate of salivary gland cells increased (all P < 0.05). Compared with other drug groups, the medium dose of Huashi Runzao could significantly activate the Hippo-YAP/TAZ pathway, improve the lymphocyte infiltration of submandibular gland tissue, reduce the water intake of mice, increase the salivary flow rate, increase the AQP5 protein expression, and reduce the apoptosis rate of salivary gland cells (all P < 0.05). in conclusion The Huashirunzao recipe can alleviate the pathological damage and apoptosis of submandibular gland cells in SS model mice and improve the overall secretory function of submandibular gland tissue, which may be related to its activation of the Hippo pathway and downregulation of YAP/TAZ levels. Objective To explore the effects of modulating the Hippo-YAP/TAZ pathway using Huashi Runzao Prescription (HRP) on the apoptosis and salivary secretion function of submandibular gland cells in a naive non-obese diabetic (NOD/Ltj) mouse model of Sjögren's syndrome (SS). Methods Eight-week-old female BALB/c mice were selected as the blank group. Eight-week-old female NOD/Ltj were randomly divided into the model, HRP-L, HRP-M, HRP-H, and hydroxychloroquine sulphate (HCQ) groups. After eight weeks of drug administration, the water consumption and salivary flow rate of each group were recorded. The histopathological damage of the submandibular gland in each group was determined using hematoxylin and eosin staining, the apoptosis rate of the submandibular gland was determined using TUNEL staining, and AQP5, Bax, Bcl-2, beclin-1, YAP, p-YAP, and TAZ expressions in submandibular gland tissues were determined using immunohistochemistry and Western blotting. Results Compared with the blank group, the Hippo-YAP/TAZ pathway was inhibited in the submandibular gland tissues of the model group, lymphocyte infiltration increased, water consumption increased, salivary flow rate decreased, AQP5 expression decreased, and submandibular gland cells apoptosis rate increased (P<0.05). Compared with all other administered groups, in the HRP-M group, the Hippo-YAZ/TAZ pathway was significantly activated, lymphocyte infiltration in submandibular gland tissues was reduced, water consumption was reduced, the salivary flow rate and AQP5 expression were increased, and the apoptosis rate of submandibular gland cells was reduced (P<0.05). Conclusion HRP reduced the pathological damage and apoptosis of submandibular gland cells in SS model mice. It improved the overall secretory function of submandibular gland tissues, which may be related to Hippo pathway activation and downregulating YAP/TAZ expression. [ABSTRACT FROM AUTHOR]

Objective: The effect of apigenin (QCS) on cardiomyocyte apoptosis in diabetic cardiomyopathy (DCM) rats by regulating Hippo-YAP signaling pathway. Methods: SD rats were divided into blank control group (CK) group (volume normal saline), Model group (volume normal saline), QCS low dose (QCS-L) group (20 mg/kg QCS), QCS high dose (QCS-H) group (40 mg/kg QCS), QCS-H+Verteporfin (Verteporfin) (Hippo-YAP signaling pathway inhibitor) group (40mg/kg QCS+10 mg/kg Verteporfin), 10 rats in each group. Cardiac function, fasting plasma glucose (FPG), fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), serum creatine kinase (CK), lactate dehydrogenase (LDH), cardiac troponin I (cTnI), superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) in myocardial tissue were detected. The morphology of myocardial tissue was detected by hematoxylin-eosin (HE) staining. The myocardial cell apoptosis was detected by TUNEL staining. The expression of B cell lymphoma-2 (Bcl-2), Yes related protein (YAP), cleaved caspase-3 (cleaved caspase-3), Bcl-2 related X protein (Bax) and transcription coactivator (TAZ) in myocardial tissue were detected by Western blot (Western blot). Results: Compared with CK group, the myocardial tissue in Model group was severely damaged, and the heart rate (HR), mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), the activity of SOD and CAT, and the expression of Bcl-2, YAP and TAZ protein were significantly decreased in model group, FPG, FINS, insulin resistance index (HOMA-IR), serum creatine kinase (CK), lactate dehydrogenase (LDH) and cardiac troponin (cTnI) levels, myocardial tissue cell apoptosis rate, MDA content, cleaved caspase-3 and Bax protein expression increased (P<0.05). Compared with Model group, the myocardial tissue morphology of rats in QCS-L group and QCS-H group were significantly improved, HR, MAP, LVSP levels, myocardial tissue SOD and CAT activity, Bcl-2, YAP, TAZ protein levels were increased, FPG, FINS, HOMA-IR levels, serum CK, LDH and cTnI levels, myocardial tissue cell apoptosis rate, MDA content, cleaved caspase-3, Bax protein expression decreased (P<0.05). Verteporfin could alleviate the improvement of QCS on cardiomyocyte apoptosis in DCM rats. Conclusion: QCS can reduce the blood glucose level and myocardial cell apoptosis rate in DCM rats, reduce the degree of oxidative stress and myocardial injury, which may be related to the activation of Hippo-YAP signaling pathway. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of baicalin (BAI) on the proliferation and apoptosis of ovarian granulosa cells in polycystic ovary syndrome (PCOS) and its mechanisms. Methods: Forty-eight female SD rats were randomly divided into the control group, PCOS group, BAI low, middle and high dose groups and positive control group, with 8 rats in each group. PCOS model was established by subcutaneous injection of DHEA, and rats in the low, medium, and high dose groups of BAI were given baicalin (12.5, 25, and 50 mg/kg) by gavage. The Positive control group was given intraperitoneal injection (10 mg/kg) of Vitipofen; The control group and PCOS group were given equal volume 0.9% NaCl solution instead. Once a day for 4 weeks. The serum sex hormone levels were detected by enzyme-linked immunosorbant assay (ELISA). The pathological changes of ovary were observed by HE staining. After successful modeling, ovarian granulosa cells were extracted, and their proliferation was detected by MTT assay. Apoptosis was detected by Annex- inV-FITC/PI double staining and TUNEL staining. The relative expressions of BCL2 and BAX, P13K/AKT pathway related proteins P13K, AKT, p-P13K and p-AKT were detected by western blot. Results: The extracted ovarian granulosa cells are fibroblast-like and adhered well, with positive expression of specific proteins FSHR. Compared with the control group, the ovarian structure of rats in the PCOS group is disrupted, with an increase in bleeding, congestion, and the number of cystic follicles, as well as thickening of the white membrane. The levels of serum testosterone (T) and luteinizing hormone (LH), the apoptosis rate of ovarian granulosa cells and the expression of Bax protein are significantly increased (P<0.05), while the levels of Estradiolm (E2) and follicle stimulating hormone (FSH), the viability of ovarian granulosa cells, the expression of Bcl-2, p-P13K, and p-AKT protein are significantly decreased (P<0.05). Compared with PCOS group, BAI groups have obvious recovery of polycystic features, significant reduction in cysts, and complete arrangement of ovarian structure and granular cell layer. The levels of E2, and FSH in serum, the viability of ovarian granulosa cells, the expressions of Bcl-2, p-P13K and p-AKT protein are significantly increased in a dose-dependent manner (P<0.05), while the levels of T and LH in serum, the apoptosis rate of ovarian granulosa cells and the expression of Bax protein are significantly decreased in a dose-dependent manner (P<0.05). Conclusion: Baicalin may promote the proliferation and inhibit the apoptosis of ovarian granulosa cell, which may alleviate PCOS by activating the P13K/AKT signaling pathway. [ABSTRACT FROM AUTHOR]

AIM: To explore the expression of calcium binding and coiled-coil domain 2（ CALCOCO2） in animal models of atrial fibrillation （AF） and its role and mechanism in reversing atrial remodeling in AF mice. METHODS: Twelve rats and 12 mice were randomly divided into the following 2 groups （n=6 each）: saline control group （saline group） and angiotensin II （Ang II）-induced AF group （Ang II group）. Western blot and immunohistochemistry （IHC） were used to detect CALCOCO2 expression in rat and mouse atrial muscle tissues. Another 24 mice were randomly divided into 4 groups（ n=6 each）: saline-oeNC, Ang II-oeNC, saline-oeCALCOCO2, and Ang II-oeCALCOCO2. An adeno-associated virus was used to induce CALCOCO2 overexpression in mouse atrial myocardium. Subsequently, transthoracic echocardiography and intracardiac electrophysiological testing were used to compare mouse cardiac function among the 4 groups. Western blot and TUNEL staining were also used to evaluate the effect of CALCOCO2 on apoptosis of cardiomyocytes in AF models. Additionally, IHC was used to assess the effect of CALCOCO2 on the levels of oxidative stress-related proteins ［NADPH oxidase 2 （NOX2） and NOX4］ and fibrosis-related proteins ［collagen type I （Col I）, connexin 40 （Cx40） and α-smooth muscle actin （α-SMA）］ in AF atrial myocardium. RESULTS: The CALCOCO2 protein level in the atrial myocardium of rats and mice was significantly decreased in Ang II group compared with saline group, as detected by Western blot and IHC （0. 19±0. 01 vs 0. 32±0. 03 for rats, 0. 37±0. 10 vs 1. 00±0. 10 for mice, P<0. 01）. Compared with Ang II-oeNC group, the mice in Ang II-oeCALCOCO2 group exhibited decreased left atrial inner diameter, shorter AF duration, and increased ejection fraction （P<0. 05）. Semi-quantitative analysis of TUNEL staining revealed a significantly decreased apoptosis rate of mouse atrial myocytes in Ang II-oeCALCOCO2 group compared with Ang II-oeNC group （0. 30±0. 06 vs 0. 61±0. 03, P<0. 01）, which was consistent with the Western blot trend of apoptosis-related proteins such as BAX （1. 94±0. 34 vs 3. 14±0. 34, P<0. 05） and cleaved caspase-3 （2. 19±0. 41 vs 3. 52±0. 55, P<0. 05）. The semiquantitative results of IHC and immunofluorescence revealed significantly increased levels of oxidative stress-related proteins （NOX2 and NOX4） and fibrosis-related proteins （Col I and α-SMA）, as well as decreased Cx40 levels, in Ang IIoeNC group compared with saline-oeNC group. However, the expression levels of these proteins were significantly reversed after CALCOCO2 overexpression （all P<0. 05）. CONCLUSION: Overexpression of CALCOCO2 reverses AF-induced electrical and structural remodeling by inhibiting the oxidative stress, apoptosis and fibrosis in mouse atrial tissues. [ABSTRACT FROM AUTHOR]

Background: Doxorubicin (DOX) is a widely used chemotherapeutic agent; nevertheless, cardiotoxicity limits its effectiveness. Orlistat (Orli) is an irreversible lipase enzyme inhibitor with poor solubility and bioavailability. Furthermore, Orli has a favorable impact on the decrease in cardiometabolic risk variables. Thus, this study aimed to investigate the novel use of Orlistat Nanocrystals (Orli-Nanocrystals) to mitigate DOX-induced cardiotoxicity and to identify probable pathways behind the cardioprotective effects. Methods: The pharmacokinetic parameters—area under % dose/g heart time curve ( A U C 0 → 4 h ), Drug targeting index (DTI), and relative targeting efficiency (RTE)—were calculated. Furthermore, experimental design mice were categorized into six groups: a (1) Normal control group, (2) Orli-Free group, (3) Orli-Nanocrystals group, (4) DOX group, (5) Orli-Free-DOX group, and (6) Orli-Nanocrystals-DOX group. All treatments were intraperitoneally injected once daily for 14 days with a single dose of DOX (15 mg/kg) on the 12th day for 4, 5, and 6 groups. Results: The pharmacokinetic parameters (Cmax, AUC) following oral administration of Orli-Nanocrystals presented a significant difference (higher values) in comparison to Orli due to the enhanced extent of the absorption of nanocrystals and, subsequently, their distribution to the heart. The study results indicated that DOX caused significant cardiotoxicity, as revealed by a remarkable rise in cardiac function biomarkers like LDH and CK-MB, which involve enzyme activities. Additionally, cardiac MDA content also increased; however, glutathione peroxidase, catalase, and superoxide dismutase activities were decreased. In the same context, DOX was found to have a remarkable downregulation in Nrf2, HO-1, Sirt-1, and Bcl2, while the upregulation of NF-κB, TNF-α, and BAX gene and protein expression occurred. Pretreatment with Orli-Nanocrystals displayed the most notable recovery of the altered immunohistochemical, histological, and biochemical characteristics as compared to the Orli-Free group. Conclusions: This work is the first investigation into the potential use of antioxidant, anti-inflammatory, and anti-apoptotic characteristics of Orli-Nanocrystals to protect against DOX-induced cardiotoxicity in vivo. [ABSTRACT FROM AUTHOR]

Vincamine, a monoterpenoid indole alkaloid with vasodilatory properties, is extracted from the leaves of Vinca minor. The present study aimed to determine the potential anticancer effects of vincamine loaded in silver nanoparticles (VCN-AgNPs) in mice with Ehrlich solid carcinoma (ESC). After tumor transplantation, the mice were divided into five groups: ESC, ESC+Cisplatin (CPN; 5 mg/kg), ESC+VCN (40 mg/kg), ESC+AgNPs (6 mg/kg), and ESC+VCN-AgNPs (20 mg/kg). The administration of VCN-AgNPs to ESC-bearing mice improved their survival rate and reduced their body weight, tumor size, and tumor weight compared to the ESC group. Furthermore, VCN-AgNPs intensified oxidative stress in tumor tissues, as evidenced by elevated levels of lipid peroxidation (LPO) and nitric oxide (NO), along with a reduction in the levels of the antioxidants investigated (GSH, GPx, GR, SOD, CAT, and TAC). Furthermore, VCN-AgNPs increased the apoptotic proteins Bax and caspase-3, decreased the anti-apoptotic protein (Bcl-2), increased the inflammatory markers TNF-α and IL-1β, and inhibited angiogenesis by lowering VEGF levels in tumor tissues, all of which led to apoptosis. Furthermore, histopathological studies showed that VCN-AgNPs suppressed the progression of Ehrlich carcinoma and induced the formation of clusters of necrotic and fragmented tumor cells. VCN-AgNPs possess cytotoxic and genotoxic effects against ESC because of their pro-oxidant, pro-apoptotic, pro-inflammatory, and antiangiogenic effects. Additionally, the combination of VCN-AgNPs was more effective and safer than chemically synthesized AgNPs, as indicated by an increase in the lifespan of animals and the total tumor inhibition index. [ABSTRACT FROM AUTHOR]

Venetoclax is a BH3 mimetic that was recently approved for the treatment of acute myeloid leukemia (AML) treatment. However, the effect of venetoclax on AML remains limited, and a novel strategy is required. Here, we demonstrate for the first time that the cytotoxic effect of venetoclax drastically increased when by combined with the naturally occurring flavonoid quercetin. Combined treatment with venetoclax and quercetin caused most of AML KG-1 cells to exhibit a condensed morphology. Cell cycle analysis revealed that the combination strongly induced cell death. Caspase inhibitor blocked this cell death, and the combination induced poly (ADP-ribose) polymerase (PARP) cleavage, indicating that apoptosis was the primary mechanism. These effects were also observed in another AML cell line Kasumi-1 but not in chronic myeloid leukemia (CML) K562 cells. Public data analysis demonstrated that B-cell/CLL lymphoma 2 (Bcl-2) expression is increased in AML cells compared to other malignant tumors, and the survival and the growth of AML cell line depends on Bcl-2. We found that quercetin increased Bcl-2-associated X protein (Bax) expression in KG-1. Our study provides a novel function for quercetin and presents a promising strategy for AML treatment using venetoclax. [ABSTRACT FROM AUTHOR]

Objective: Previously, we have successfully purified and synthesized viscolin, an agent derived from Viscum coloratum extract, which has shown significant potential in the treatment of stroke. Our study aimed to evaluate the neuroprotective effects of viscolin. Methods: We first assessed the cytotoxicity of viscolin on primary neuronal cultures and determined its antioxidant and radical scavenging properties. Subsequently, we identified the optimal dose-response of viscolin in protecting against glutamate-induced neurotoxicity. Results: Our results demonstrated that viscolin at a concentration of 10 μM effectively reduced neuronal cell death up to 6 hours after glutamate-induced neurotoxicity. Additionally, we investigated the therapeutic window of opportunity and the potential of viscolin in preventing necrotic and apoptotic damage in cultured neurons exposed to oxygen glucose deprivation-induced neurotoxicity. Our findings showed that viscolin treatment significantly reduced DNA breakage, prevented the release of cytochrome c from mitochondria to cytosol, increased the expression of anti-apoptotic protein Bcl-2, decreased the expression of pro-apoptotic protein Bax, and reduced the number of TUNEL-positive cells. Additionally, our in vivo investigation demonstrated a reduction in brain infarction following middle cerebral artery occlusion. Conclusion: Viscolin has potential utility as a therapeutic agent in the treatment of stroke. [ABSTRACT FROM AUTHOR]

We aimed to investigate whether pulsed magnetic fields (PMFs) (1 mT) may have preventive effects on myocardial damage and apoptosis in rats with sepsis. Twenty-eight adult Wistar albino rats were evenly distributed among four experimental groups, each consisting of seven rats: SH, LF-PMF, HF-PMF, and CLP. Sepsis induction was carried out via the cecal ligation and puncture (CLP) method, while rats in the LF-PMF and HF-PMF groups were exposed to 7.5 Hz and 15 Hz PMF, respectively, for duration of 24 hours. Following the removal of heart tissue, histological techniques were employed for the analysis. Histological scoring of apoptosis-related Bax, Bcl-2, and Acas-3 proteins as well as cTnI were performed in the heart tissue. The myocardial damage score significantly increased in the CLP group compared to the SH group (p < 0.05). Significant decreases were observed in Bcl-2 and cTnI protein levels in the CLP group, while significant increases were detected in the PMF groups (p < 0.05). An increase in Bax and Acas-3 protein levels, as well as the Bax/Bcl-2 ratio, was observed in the CLP group, with a decrease in the PMF groups (p < 0.05). The results demonstrate that PMF application has anti-apoptotic and therapeutic effects on septic heart tissue damage. [ABSTRACT FROM AUTHOR]

Guijiajiao-Lujiaojiao (GL) is a combination of Traditional Chinese Medicine (TCM) that can be used to treat oligoasthenozoospermia (OAS). However, its mechanistic role in OAS needs to be better understood and necessitates more studies. This study was planned to investigate GL's therapeutic effects and its mechanistic role in the tripterygium wilfordii polyglycoside (GTW)-induced OAS rat model. In total, 60 Sprague–Dawley (SD) rats at 8 weeks of age were assigned to six groups: blank (NC), model (GTW), GL low-dose (GL-L, 0.3 g/kg/day), GL medium-dose (GL-M, 0.6 g/kg/day), GL high-dose (GL-H, 1.2 g/kg/day), and GL high-dose + PI3K inhibitor LY294002 (GL-H 1.2 g/kg/day + LY 1.2 mg/kg/day) groups. The model was characterized after 8 weeks to examine sperm concentration and viability, serum hormone levels, testes histopathology, and specific protein markers. The treatment efficacy was evaluated by mRNA and protein expression levels, among other parameters. Compared with the GTW group, the viability and concentration of rat spermatozoa were significantly increased after GL intervention (p <.01). Meanwhile, the serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and T hormones in rats in the GL-M and GL-H groups were significantly higher than those in the GTW group (p <.05). Furthermore, GL enhanced the proliferation of spermatogenic cells by modulating the PI3K/AKT signaling pathway, increasing and decreasing the levels of Bcl-2 and Bax proteins, respectively. It is concluded that the mechanism by which GL effectively enhanced the spermatogenic function of the GTW-induced OAS model may be attributed to the PI3K/AKT signaling pathway activation and the elevation of serum LH, FSH, and T hormone levels. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate whether the damage to male offspring induced by cadmium (Cd) exposure during embryonic period leads to the apoptosis of ovarian granulosa cells (OGCs) in the next generation of female offspring, and whether this apoptosis in the offspring was due to paternal genetic effects. Pregnant Sprague–Dawley (SD) rats were exposed to CdCl2 (0, 0.5, 2.0, or 8.0 mg/kg) by gavage daily for 20 days to produce the filial 1 (F1) generation. F1 males were mated with newly purchased females to produce the F2 generation, and the F3 generation was generated in the same way. No apoptotic bodies were observed in the OGCs of either the F2 or F3 generation as shown by electron microscopy, and a reduced OGC apoptosis rate (detected by flow cytometry) was observed in F2 OGCs from the Cd‐exposed group. Moreover, the mRNA (qRT‐PCR) levels of Bax and Bcl‐2 and the protein (western blotting) level of pro‐caspase‐8 increased in the F2 generation (p < 0.05). The expression of apoptosis‐related miRNAs (qRT‐PCR) and methylation of apoptosis‐related genes (determined via bisulfite‐sequencing PCR) in OGCs were further determined. Compared with those of the controls, the expression patterns of microRNAs (miRNAs) in the F2 offspring were different in the Cd‐exposed group. The miR‐92a‐2‐5p expression levels were decreased in both the F2 and F3 generations (p < 0.05), while the average methylation level of apoptosis‐related genes did not change significantly (except for individual loci). In summary, this study showed that the paternal genetic intergenerational effect of male Cd exposure during embryonic period induced apoptosis of OGCs in the offspring was weakened, and the transgenerational effect disappeared; nevertheless, intergenerational and transgenerational changes in apoptosis‐related genes, epigenetic modifications, DNA methylation, and miRNAs were observed, and may be important for understanding the homeostatic mechanisms of the body to alleviate the intergenerational transmission of Cd‐induced damage. [ABSTRACT FROM AUTHOR]

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BACKGROUND: Formononetin is an isoflavonoid compound widely found in red clover, astragalus, and chickweed, which has the ability to inhibit oxidative stress, inflammatory factor release, and apoptosis. OBJECTIVE: To investigate the effect of formononetin on interleukin-1β-induced chondrocyte injury and its mechanism. METHODS: (1) Cartilage tissues from patients with osteoarthritis and patients with simple meniscus injury were collected, and real-time quantitative PCR was used to detect miR-135b-5p expression. (2) Human chondrocytes were cultured in vitro, and then divided them into nine groups: cells in normal control group were cultured for 48 hours with no treatment; cells in interleukin-1β group were treated with interleukin-1β for 48 hours; cells in interleukin-1β+lowdose formononetin group were treated with 25 μmol/L formononetin for 24 hours followed by treatment with interleukin-1β for 48 hours; cells in interleukin1β+middle-dose formononetin group were treated with 50 μmol/L formononetin for 24 hours followed by treatment with interleukin-1β for 48 hours; cells in interleukin-1β+high-dose formononetin group were treated with 100 μmol/L formononetin for 24 hours followed by treatment with interleukin-1β for 48 hours; cells in interleukin-1β+miR NC group were treated with miR NC for 6 hours followed by treatment with interleukin-1β for 48 hours; cells in interleukin1β+miR-135b-5p group were treated with miR-135b-5p mimics for 6 hours followed by treatment with interleukin-1β for 48 hours; cells in interleukin-1β+highdose formononetin+anti-miR-NC group were treated with anti-miR-NC for 6 hours, then treated with 100 μmol/L formononetin for 24 hours, and finally treated with interleukin-1β for 48 hours; cells in interleukin-1β+high-dose formononetin+anti-miR-135b-5p group were treated with anti-miR-135b-5p for 6 hours, then treated with 100 μmol/L formononetin for 24 hours, and finally treated with interleukin-1β for 48 hours. Relevant tests are performed after treatment. RESULTS AND CONCLUSION: The expression level of miR-135b-5p in cartilage tissue of patients with osteoarthritis was significantly lower than that of patients with simple meniscus injury (P < 0.05). Compared with the normal control group, the expression level of miR-135b-5p, proliferative ability, activity of superoxide dismutase, and expression levels of collagenase II protein and Bcl-2 protein in chondrocytes were lower in the interleukin-1β group (P < 0.05), while apoptotic rate, lactate dehydrogenase activity, malondialdehyde level, levels of proinflammatory factors, and expression levels of matrix metalloproteinase-13 protein and Bax protein were higher in the interleukin-1β group (P < 0.05). Formononetin inhibited chondrocyte damage caused by interleukin-1β in a concentrationdependent manner. Transfection of miR-135b-5p mimics elevated miR-135b-5p expression in the interleukin-1β group and inhibited chondrocyte damage induced by interleukin-1β; transfection of anti-miR-135b-5p decreased miR-135b-5p expression in the interleukin-1β+high-dose formononetin group and inhibited the effect of formononetin on chondrocytes. To conclude, the protective effect of formononetin on chondrocyte injury induced by interleukin-1β may be related to the regulation of miR-135b-5p expression. [ABSTRACT FROM AUTHOR]

BACKGROUND: Chondrocyte apoptosis is an important factor in the development of osteoarthritis, and Complanatoside A has a flavonoid effect, which can inhibit apoptosis of various cells, but its effect on chondrocyte apoptosis and the mechanism of action are not clear. OBJECTIVE: To investigate the intrinsic association and mechanism of Complanatoside A in chondrocyte apoptosis based on the Wnt/β-catenin signaling pathway. METHODS: (1) The cartilage tissues of the femur and tibia transected during knee arthroplasty were collected, and chondrocytes were isolated, cultured in vitro, and identified. (2) Cell counting kit-8 was used to detect the optimal intervention concentration of Complanatoside A in the concentration range of 0- 160 μmol/L. (3) Chondrocytes were divided into blank group, sodium nitroprusside (1.5 mmol/L)-induced group, and sodium nitroprusside (1.5 mmol/L)+ Complanatoside A (5 μmol/L) group. The viability and apoptosis rate of the cells in each group were detected by cell counting kit-8 and flow cytometry. The expression of type II collagen and SOX9 was detected by immunofluorescence staining. The expression of apoptosis-related proteins and Wnt/β-catenin pathway proteins was detected by western blot assay. RESULTS AND CONCLUSION: The cells extracted in vitro were cultured and stained, and were clearly identified as chondrocytes. Complanatoside A had no obvious cytotoxicity to chondrocytes in the concentration range of 0-80 μmol/L, and significantly improved the chondrocyte viability in the concentration range of 2.5-10 μmol/L, especially when the concentration was 5 μmol/L. The apoptotic rate of chondrocytes was higher in the sodium nitroprusside-induced group than the blank control group, while the apoptotic rate was lower in the sodium nitroprusside+Complanatoside A group than the sodium nitroprusside-induced group. The fluorescence intensity of type II collagen and SOX9 in chondrocytes was weaker in the sodium nitroprusside-induced group than the blank control group, while the fluorescence intensity of type II collagen and SOX9 in the sodium nitroprusside+Complanatoside A group was higher than that of the sodium nitroprusside-induced group. In the sodium nitroprusside-induced group, the protein expression of Bax, Caspase-3, matrix metalloproteinase 13, Wnt3a, Wnt5a and β-catenin was higher than that of the blank control group, while the protein expression of Bcl-2 was lower than that of the blank control group. In the sodium nitroprusside+Complanatoside A group, except for the protein expression of Bcl-2 which was higher than that of the sodium nitroprusside-induced group, the expression of the other aforementioned proteins was lower than that of the sodium nitroprusside-induced group. To conclude, Complanatoside A has a certain inhibitory effect on chondrocyte apoptosis, which could regulate apoptosis-related proteins and promote the expression of chondrocyte regulatory factors, and presumably might play a role through inhibiting the Wnt/β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]

BACKGROUND: Previous studies have confirmed that Wen-Shen-Tong-Du Decoction can promote the recovery of spinal cord injury by inhibiting pyroptosis of splenic B cells, promoting the phagocytosis of myelin debris by microvascular endothelial cells, affecting the migration and infiltration of microglia, promoting the recovery of damaged neurons, and decreasing neuronal apoptosis after spinal cord injury, but the mechanism of this is still not clear. OBJECTIVE: To investigate the effect of Wen-Shen-Tong-Du Decoction on the triggering receptor expressed on myeloid cells 2 (TREM2) and PI3K/Akt signaling pathways in mice following spinal cord injury. METHODS: Thirty-six C57BL/6 mice were selected and randomly divided into a sham-operation group, a model group and a Wen-Shen-Tong-Du Decoction group, with 12 mice in each group. In the model and Wen-Shen-Tong-Du Decoction groups, mouse models of T10 spinal cord injury were prepared by the modified Allen’s method. On the 1st day after modeling, the Wen-Shen-Tong-Du Decoction group was given Wen-Shen-Tong-Du Decoction by gavage, and the sham-operation group and the model group were given saline by gavage once a day for 28 days. During the drug administration period, mouse motor function was evaluated by Basso Mouse Scale score and inclined plane test. On the 7th and 28th days after modeling, hematoxylin-eosin staining was used to observe the histopathological changes in the spinal cord tissue of the mice; immunofluorescence double staining was used to detect the protein expression of ionized calcium binding adaptor molecule 1 (IBA1) and TREM2; and western blot assay was used to detect the expression of TREM2, PI3K, p-PI3K, Akt, p-Akt, Bcl2, Bax and Caspase3 in spinal cord tissue. RESULTS AND CONCLUSION: Basso Mouse Scale scores and inclined plane test results indicated that the motor function of the mouse hindlimbs was declined after spinal cord injury, and Wen-Shen-Tong-Du Decoction significantly improved motor function in mice with spinal cord injury. Hematoxylin-eosin staining results revealed that Wen-Shen-Tong-Du Decoction significantly ameliorated the pathological structure of spinal cord tissue compared with the model group, manifesting as reduced degrees of dorsal white matter and neuronal atrophy, decreased cytoplasmic vacuolization, and reduced inflammatory cell infiltration. Immunofluorescence double staining results showed that on the 7th day after modeling, the protein expression of IBA1 and TREM2 in the model group was lower than that in the sham-operation group (P < 0.05), and the protein expression of IBA1 and TREM2 in the Wen-Shen-Tong-Du Decoction group was higher than that in the model group (P < 0.05); on the 28th day after modeling, the protein expression of TREM2 in the model group was lower than that in the sham-operation group (P < 0.05), and the protein expression of TREM2 in the spinal cord tissue of the mice in the Wen-Shen-Tong-Du Decoction group was higher than that in the model group (P < 0.05). Western blot results analysis demonstrated that on the 7th day after modeling, compared with the sham-operation group, the model group exhibited a significant reduction in TREM2, PI3K, and Bcl2/Bax (P < 0.05), as well as a significant increase in p-Akt, Bax and p-Akt/Aktp-PI3K (P < 0.05); compared with the model group, the Wen-Shen-Tong-Du Decoction group showed a significant increase in TREM2, PI3K, p-PI3K, Akt, p-Akt, Bcl2, p-PI3K/PI3K, p-Akt/Ak, and Bcl2/Bax (P < 0.05), as well as a significant decrease in Bax and Caspase3 protein expression (P < 0.05). On the 28th day after modeling, compared with the sham-operation group, the model group exhibited a significant reduction in TREM2, PI3K, p-PI3K, Akt, p-Akt, Bcl2 and Bcl2/Bax (P < 0.05), as well as a significant increase in Bax protein expression (P < 0.05); compared with the model group, the Wen-Shen-Tong-Du Decoction group showed a significant increase in TREM2, PI3K, Akt, p-Akt, Bcl2, and Bcl2/Bax (P < 0.05), as well as a significant decrease in Bax protein expression (P < 0.05). To conclude, Wen-Shen-Tong-Du Decoction may activate the PI3K/Akt signaling pathway by up-regulating the expression of TREM2 protein in microglia, and then inhibit neuronal apoptosis, thus exerting neuroprotective effects and promoting the repair of spinal cord injury. [ABSTRACT FROM AUTHOR]

Objective: We investigated the mechanism of Dexmedetomidine (Dex) in infant rats with brain injury. Methods: The infant rats underwent brain injury modelling. The motor function, spatial learning and memory abilities in rats, and the hippocampal CA1 region Nissl body level and apoptosis were evaluated by behavioural tests and histological stainings. Levels of the hippocampal CA1 region p-IRE1α, nuclear/cytoplasmic p65, CHOP, Bax and Bcl-2 proteins were determined by Western blot. Results: Propofol anaesthesia caused brain injury in infant rats. Dex increased the hippocampal CA1 region Nissl body level, abated cell apoptosis, reduced p-IRE1α, ATF6, p-PERK/PERK and CHOP levels, decreased the Bax protein level, elevated the Bcl-2 protein level, and alleviated brain injury in infant rats. After ERS induction and the NF-κB pathway inhibition, the hippocampal CA1 region nuclear/cytoplasmic p65 ratio, CHOP level, and apoptosis were reduced in infant rats with brain injury treated with Dex, while the learning and memory abilities of rats were enhanced. Conclusion: Dex reduced the hippocampal CA1 region cell apoptosis and enhanced learning and memory abilities by inhibiting the ERS-mediated IRE1α/NF-κB/CHOP pathway, thereby alleviating brain injury in infant rats. [ABSTRACT FROM AUTHOR]

Objective(s): Trans-sodium crocetinate (TSC) is one of the crocetin derivations that is more soluble and stable than crocetin and its cis form. It easily crosses the blood-brain barrier. TSC has neuroprotective effects. Bisphenol A (BPA) is an endocrine-mimicking compound that induces Parkinson-like disease by impacting the dopaminergic system. In this research, the effects of TSCs on BPA-induced Parkinsonlike symptoms via behavioral and molecular assays have been investigated. Materials and Methods: Male Wistar rats received BPA (75 mg/kg, gavage), TSC (10, 20, and 40 mg/kg), and levodopa (L-dopa) (10 mg/kg) via intraperitoneal injection (IP) for 28 days. Parkinsonian-like motor features were evaluated using bar test, rotarod, and open field experiments. Malondialdehyde (MDA) and glutathione (GSH) levels were also measured as the most important indicators of oxidative stress. Western blotting was performed for the molecular assays of alpha-synuclein (a-syn), Bcl-2, Bax, caspase-3, Beclin, and LC3 I/II proteins. Results: Our analyses indicated that treatment with TSC at high dose reduces MDA levels and protects GSH reserves. TSC can also increase anti-apoptotic Bcl-2 and decrease pro-apoptotic Bax and caspase-3 proteins. While it does not affect autophagy markers, TSC decreased a-syn protein expression, reduced the catalepsy time, and improved the time spent staying on the rotating bar and the locomotor activity. Conclusion: Overall, TSC likely ameliorates BPA-mediated Parkinson' s-like symptoms by suppressing oxidative stress inhibition. This leads to reduced a-syn expression, which ultimately results in apoptosis inductions. Therefore, TSC can serve as a promising exploratory target for future research aimed at controlling Parkinson's disease. [ABSTRACT FROM AUTHOR]

Background: Oral mucositis (OM) is an acute debilitating side effects of chemoradiotherapy (CRT) in head and neck cancer (HNC) patients. Salivary biomarkers are noninvasive and aid in early diagnosis and disease prevention. Objective: This study analyzed Bcl-2, Mcl-1, and Bax as potential biomarkers in CRT-induced OM in HNC patients. Methods: Forty-six HNC patients receiving CRT with OM were evaluated using WHO grading. Two time points were observed for saliva sample collection: Day 0 (prior to CRT) and Day 30 (CRT with OM) and from 46 healthy volunteers. Using an enzyme-linked immunosorbent assay (ELISA), levels of Bcl-2, Mcl-1, and Bax were measured. Oral mucositis progression was analyzed using Friedman’s test, and statistical significance between means of different salivary marker was determined using the ANOVA test. Results: From Day 0 to Day 30, grades of OM were statistically significant (P < 0.05). Apoptotic biomarkers showed statistically significant higher expression of proapoptotic regulator Bax and lower expressions of antiapoptotic markers Bcl-2 and Mcl-1 on Day 30 (severe OM) compared to Day 0 and healthy controls (P < 0.0001). Conclusion: Our research showcased Bax, Bcl-2, and Mcl-1 as early predictive indicators of OM, and targeted drugs should be developed against these to prevent onset and severity of oral mucositis that improves patients quality of life.

The most common mutation in southern Chinese individuals with late-onset multiple acyl-coenzyme A dehydrogenase deficiency (MADD; a fatty acid metabolism disorder) is c.250G > A (p.Ala84Thr) in the electron transfer flavoprotein dehydrogenase gene (ETFDH). Various phenotypes, including episodic weakness or rhabdomyolysis, exercise intolerance, and peripheral neuropathy, have been reported in both muscular and neuronal contexts. Our cellular models of MADD exhibit neurite growth defects and excessive apoptosis. Given that axonal degeneration and neuronal apoptosis may be regulated by B-cell lymphoma (BCL)-2 family proteins and mitochondrial outer membrane permeabilization through the activation of proapoptotic molecules, we measured the expression levels of proapoptotic BCL-2 family proteins (e.g., BCL-2-associated X protein and p53-upregulated modulator of apoptosis), cytochrome c, caspase-3, and caspase-9 in NSC-34 cells carrying the most common ETFDH mutation. The levels of these proteins were higher in the mutant cells than in the wide-type cells. Subsequent treatment of the mutant cells with coenzyme Q10 downregulated activated protein expression and mitigated neurite growth defects. These results suggest that the activation of the BCL-2/mitochondrial outer membrane permeabilization/apoptosis pathway promotes apoptosis in cellular models of MADD and that coenzyme Q10 can reverse this effect. Our findings aid the development of novel therapeutic strategies for reducing axonal degeneration and neuronal apoptosis in MADD. [ABSTRACT FROM AUTHOR]

Introduction: Pituitary adenomas (PAs) are benign tumors with high prevalence and, occasionally, aggressive course. The tumorigenesis of these lesions is not completely understood at the molecular level. BAK1 and BAX proteins play fundamental roles in apoptosis and seem to interact with VDAC proteins, whose expressions have been markedly altered in cancer, impacting their prognosis. Objective: to evaluate the gene expression of VDAC1, VDAC2, BAK1 and BAX and their association with clinical and imaging characteristics in PA. Methods: Clinical-epidemiological data were collected from 117 tumor samples from patients affected by PA. Invasiveness was assessed by the Knosp scale. Gene expression was examined by real-time PCR. Relative expression analysis was performed by 2^(-DDCt) method. Results: The sample was mainly composed of women (69/117 -- 57.2%). Tumor subtypes observed were Non-Functioning (NF) (73/117 -- 62.4%), Acromegaly (24/117 -- 20.5%) and Cushing's Disease (CD) (20/117 -- 17.1%). Compared to normal tissue, there was a significant reduction in VDAC1 expression in the Acromegaly (p=0.029) and NF (p=0.002) groups. BAX expression was lower in all groups (p <0.001; p=0.007; P =0.005). No difference was found in VDAC2 and BAK1 expression, compared to normal pituitary. Overexpression of VDAC2 occurred in PAs with post-surgical regrowth (p=0.042). A strongly negative correlation was observed in BAX and BAK1 expression in CD. Conclusion: The results indicated that downregulations of VDAC1 and BAX may be related to resistance to apoptosis. In contrast, overexpression of VDAC2 in regrowing PAs suggests an antiapoptotic role for this gene. In summary, the genes evaluated might be involved in the biopathology of PAs. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of sodium glucose cotransporter 2 inhibitor (SGLT-2i) regulating the TLRs/MyD88 pathway on high glucose induced autophagy and apoptosis in human aortic endothelial cells (HAECs). Methods: Using HAECs, the cells were divided into NG group (5.5 mmol/L glucose), HG group (30 mmol/L glucose), and SGLT-2i+HG group (30 mmol/L glucose+50) μ Mol/L of SGLT-2 inhibitor. Detect the growth vitality; the level of cell apoptosis; the expression levels of TLRs and MyD88 mRNA; the levels of TLRs, MyD88, Bax, Bcl-2, Beclin 1, and LC3II in each group of cells. Results: Compared with the NG group, the cell proliferation rate of the HG group was significantly reduced (P<0.05); Compared with the HG group, the SGLT-2i+HG group showed a significant increase in cell proliferation rate (P<0.05). At 12 hours and 24 hours, compared with the NG group, the apoptosis rate of cells in the HG group was significantly increased (P<0.05); Compared with the HG group, the apoptosis rate of SGLT-2i+HG group was significantly reduced (P<0.05). Compared with the NG group, the expression levels of apoptosis related proteins Bax and Bcl-2 in the HG group increased (P<0.05); Compared with the HG group, the expression levels of Bax and Bcl-2 in cells of the SGLT-2i+HG group decreased (P<0.05). Compared with the NG group, the levels of autophagy related proteins Beclin 1 and LC3II in the HG group increased (P<0.05); Compared with the HG group, the levels of Beclin 1 and LC3II in cells of the SGLT-2i+HG group decreased (P<0.05). Compared with the NG group, the expression levels of TLRs and MyD88 proteins in the HG group increased (P<0.05); Compared with the HG group, the expression levels of TLRs and MyD88 in the SGLT-2i+HG group decreased (P<0.05). Conclusion: SGLT-2 inhibits high glucose induced autophagy and apoptosis in human aortic endothelial cells by regulating the TLRsMyD88 pathway. [ABSTRACT FROM AUTHOR]

Difenoconazole (DIF) is a fungicide used to control various fungi. It is absorbed on the surface of different plants and contributes significantly to increased crop production. However, DIF is reported to exhibit toxicity to fungi and to aquatic plants, fish, and mammals, including humans, causing adverse effects. However, research on the impact of DIF on the mammary epithelial cells of herbivorous bovines is limited. DIF-induced damage and accumulation in the mammary glands can have direct and indirect effects on humans. Therefore, we investigated the effects and mechanisms of DIF toxicity in MAC-T cells. The current study revealed that DIF reduces cell viability and proliferation while triggering apoptotic cell death through the upregulation of pro-apoptotic proteins, including cleaved caspase 3 and Bcl-2-associated X protein (BAX), and the downregulation of leukemia type 2 (BCL-2). DIF also induced endoplasmic reticulum (ER) stress by increasing the expression of genes or proteins of Bip/GRP78, protein disulfide isomerase (PDI), activating transcription factor 4 (ATF4), C/EBP homologous protein (CHOP), and endoplasmic reticulum oxidoreductase 1 Alpha (ERO1-Lα). We demonstrated that DIF induces mitochondria-mediated apoptosis in MAC-T cells by activating ER stress pathways. This cellular damage resulted in a significant increase in the expression of inflammatory response genes and proteins, including cyclooxygenase 2 (COX2), transforming growth factor beta 3 (TGFB3), CCAAT enhancer binding protein delta (CEBPD), and iNOS, in DIF-treated groups. In addition, spheroid formation by MAC-T cells was suppressed by DIF treatment. Our findings suggest that DIF exposure in dairy cows may harm mammary gland function and health and may indirectly affect human consumption of milk. [ABSTRACT FROM AUTHOR]

Microgravity can induce alterations in liver morphology, structure, and function, with mitochondria playing an important role in these changes. Tail suspension (TS) is a well-established model for simulating the effects of microgravity on muscles and bones, but its impact on liver function remains unclear. In the current study, we explored the regulatory mechanisms of apoptosis, autophagy, fission, and fusion in maintaining liver mitochondrial homeostasis in mice subjected to TS for 2 or 4 weeks (TS2 and TS4). The results showed the following: (1) No significant differences were observed in nuclear ultrastructure or DNA fragmentation between the control and TS-treated groups. (2) No significant differences were detected in the mitochondrial area ratio among the three groups. (3) Cysteine aspartic acid-specific protease 3 (Caspase3) activity and the Bcl-2-associated X protein (bax)/B-cell lymphoma-2 (bcl2) ratio were not higher in the TS2 and TS4 groups compared to the control group. (4) dynamin-related protein 1 (DRP1) protein expression was increased, while mitochondrial fission factor (MFF) protein levels were decreased in the TS2 and TS4 groups compared to the control, suggesting stable mitochondrial fission. (5) No significant differences were observed in the optic atrophy 1 (OPA1), mitofusin 1 and 2 (MFN1 and MFN2) protein expression levels across the three groups. (6) Mitochondrial autophagy vesicles were present in the TS2 and TS4 groups, with a significant increase in Parkin phosphorylation corresponding to the duration of the TS treatment. (7) ATP synthase and citrate synthase activities were significantly elevated in the TS2 group compared to the control group but were significantly reduced in the TS4 group compared to the TS2 group. In summary, the coordinated regulation of apoptosis, mitochondrial fission and fusion, and particularly mitochondrial autophagy preserved mitochondrial morphology and contributed to the restoration of the activities of these two key mitochondrial enzymes, thereby maintaining liver mitochondrial homeostasis in mice under TS conditions. [ABSTRACT FROM AUTHOR]

5-Demethylnobiletin is the active ingredient in citrus polymethoxyflavones that could inhibit the proliferation of several tumor cells. However, the anti-tumor effect of 5-Demethylnobiletin on glioblastoma and the underlying molecular mechanisms are remains unknown. In our study, 5-Demethylnobiletin markedly inhibited the viability, migration and invasion of glioblastoma U87-MG, A172 and U251 cells. Further research revealed that 5-Demethylnobiletin induces cell cycle arrest at the G0/G1 phase in glioblastoma cells by downregulating Cyclin D1 and CDK6 expression levels. Furthermore, 5-Demethylnobiletin significantly induced glioblastoma cells apoptosis by upregulating the protein levels of Bax and downregulating the protein level of Bcl-2, subsequently increasing the expression of cleaved caspase-3 and cleaved caspase-9. Mechanically, 5-Demethylnobiletin trigged G0/G1 phase arrest and apoptosis by inhibiting the ERK1/2, AKT and STAT3 signaling pathway. Furthermore, 5-Demethylnobiletin inhibition of U87-MG cell growth was reproducible in vivo model. Therefore, 5-Demethylnobiletin is a promising bioactive agent that might be used as glioblastoma treatment drug. [ABSTRACT FROM AUTHOR]

Objective To analyze the effects of agatroban combined with mitochondrial transplantation on endothelial function and hemorheology in mice with cerebral ischemia-reperfusion injury. Methods 50 SPF grade Kunming mice, with 10 as the normal group, 38 mices were successfully modeled and divided into a model group of 9, a mitochondrial transplantation group of 9, an Agatroban group of 10, and a combination group of 10. The mitochondrial transplantation group was given exogenous mitochondrial extract, the agatroban group was given agatroban, the combined group was given agatroban combined with mitochondrial transplantation, and the blank group and model group were injected with normal saline. Observation of changes in cerebral infarction volume, pathological morphology of brain tissue, endothelial function, hemorheology, and relative expression of apoptosis related proteins in mice after 7 days of intervention. Results Compared with normal group, cerebral infarction volume ratio, ET-1, whole blood viscosity, plasma viscosity, Caspase-3 and Bax protein expression levels were increased in model group, mitochondrial transplantation group, agatroban group and combined group, while NO level and Bcl-2 protein expression levels were decreased (P < 0.05). Compared with the model group, the cerebral infarction volume ratio, ET-1, whole blood viscosity, plasma viscosity, Caspase-3 and Bax protein expressions in the mitochondrial transplantation group, agatroban group and combined group were decreased, and the combined group was lower than the mitochondrial transplantation group and agatroban group, and the NO level and Bcl-2 protein expression were increased. The combined group was higher than that of mitochondrial transplantation group and agatriban group (P < 0.05). Conclusion In mice with cerebral ischemia-reperfusion injury, endothelial function injury was improved, hemorheology was regulated, and brain injury was alleviated after agatroban combined with mitochondrial transplantation. [ABSTRACT FROM AUTHOR]

Lung ischemia-reperfusion injury (LIRI) causes oxidative stress, inflammation, and immune system activation. The Nrf2/Keap1/HO-1 pathway is important in cellular defense against these effects. Quercetin, a flavonoid with antioxidant, anti-inflammatory, and anti-cancer properties, has been investigated. Our aim in this study was to investigate the effect of quercetin on preventing lung ischemia-reperfusion injury and the role of the Nrf2/Keap1/HO-1 pathway. Sixty-four male Wistar rats were divided into four distinct groups(n = 16). Sham, lung ischemia-reperfusion (LIR), Saline + LIR, Quercetin + LIR (30 mg/kg i.p for a week before LIR). LIR groups were subjected to 60 min of ischemia (left pulmonary artery, vein, and bronchus) and 120 min of reperfusion. Our assessment encompassed a comprehensive analysis of various factors, including the evaluation of expression Nrf2, Keap1, and Heme Oxygenase-1 (HO-1) levels and NF-κB protein. Furthermore, we examined markers related to inflammation (interleukin-1β and tumor necrosis factor alpha), oxidative stress (malondialdehyde, total oxidant status, superoxide dismutase, glutathione peroxidase, total antioxidant capacity), lung edema (Wet/dry lung weight ratio and total protein concentration), apoptosis (Bax and Bcl2 protein), and histopathological alterations (intra-alveolar edema, alveolar hemorrhage, and neutrophil infiltration). Our results show that ischemia-reperfusion results in heightened inflammation, oxidative stress, apoptosis, lung edema, and histopathological damage. Quercetin showed preventive effects by reducing these markers, acting through modulation of the Nrf2/Keap1 pathway and inhibiting the NF-κB pathway. This anti-inflammatory effect, complementary to the antioxidant effects of quercetin, provides a multifaceted approach to cell protection that is important for developing therapeutic strategies against ischemia-reperfusion injury and could be helpful in preventive strategies against ischemia-reperfusion. [ABSTRACT FROM AUTHOR]

AIM: To investigate the effect of protein kinase C （PKC）-mammalian target of rapamycin （mTOR） pathway on the apoptosis of gastric smooth muscle cells in diabetic rats, and to explore the related mechanism. METHODS: Male and female 4-week-old specific pathogen-free Sprague-Dawley rats were randomly allocated to control, diabetes, diabetes+0. 1 μmol/L phorbol 12, 13-myristate acetate （PMA）, diabetes+0. 2 μmol/L PMA, diabetes+0. 5 μmol/L PMA, diabetes+2 μmol/L bisindolylmaleimide I （GF109203X）, diabetes+5 μmol/L GF109203X, and diabetes+ 10 μmol/L GF109203X groups （n=9）. Primary rat gastric smooth muscle cells cultured in a high-glucose environment in vitro were treated with 0. 1 μmol/L PMA, 0. 2 μmol/L PMA, 0. 5 μmol/L PMA, 2 μmol/L GF109203X, 5 μmol/L GF109203X, 10 μmol/L GF109203X, or vehicle. The isolated muscle technique was used to evaluate the spontaneous contraction of gastric antral smooth muscle. The pathologic changes and apoptosis of rat gastric smooth muscle were identified using HE staining and flow cytometry, respectively. The expression levels of stem cell factor （SCF）, c-Kit, mTOR, phosphorylated （p）-mTOR, p70 ribosomal protein S6 kinase（ p70S6K）, p-p70S6K, eIF4E-binding protein 1（ 4E-BP1）, p-4E-BP1, caspase-3, B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax） in gastric smooth muscle were measured by Western blot. RESULTS: The contractility of the gastric antral smooth muscle was lower （P<0. 05）, gastric smooth muscle atrophy and the apoptosis rate was higher （P<0. 01）, and the expression of SCF, c-Kit, and p-mTOR was lower（ P<0. 05 or P<0. 01） in the diabetes group than in the control group. There was no significant difference in the spontaneous contraction of the gastric antral smooth muscle between the diabetes and diabetes+0. 1 μmol/L PMA groups, but it was significantly greater in the diabetes+0. 2 μmol/L PMA and diabetes+0. 5 μmol/L PMA groups （P<0. 05 or P<0. 01）. The phosphorylation levels of mTOR, p70S6K （T421/S424）, and p70S6K （T389） were also higher in the diabetes + 0. 2 μmol/L PMA and diabetes+0. 5 μmol/L PMA groups than in diabetes group （P<0. 01）. Furthermore, the phosphorylation of 4E-BP1 was higher（ P<0. 05） in the diabetes+0. 5 μmol/L PMA group than in the diabetes group（ P<0. 01）. There was no significant difference in the spontaneous contraction of the gastric antral smooth muscle between the diabetes and diabetes+2 μmol/L GF109203X groups, but it was significantly lower in the diabetes+5 μmol/L GF109203X and diabetes+10 μmol/L GF109203X groups （P<0. 05 or P<0. 01）. The phosphorylation levels of mTOR and p70S6K （T389） were lower in the diabetes+5 μmol/L GF109203X and diabetes+10 μmol/L GF109203X groups （P<0. 05）, the phosphorylation of p70S6K（ T421/S424） was lower in the diabetes+10 μmol/L GF109203X group（ P<0. 05）, and the phosphorylation of 4EBP1 was lower in the diabetes+GF109203X （2, 5 and 10 μmol/L） groups （P<0. 05） than in the diabetes group. Compared with the CK group, the apoptosis rate was high in the GF109203X-treated（ 2, 5 and 10 μmol/L） groups（ P<0. 01）, the expression of Bax and caspase-3 was low （P<0. 05）, and the expression of Bcl-2 was high in the PMA-treated （0. 1, 0. 2 and 0. 5 μmol/L） groups（ P<0. 05）. CONCLUSION: The downregulation of the PKC-mTOR pathway in the gastric smooth muscle of diabetic rats leads to apoptosis of gastric smooth muscle cells through Bax/Bcl-2 and caspase-3 and the downregulation of SCF and c-Kit, which may play an important role in the gastric dysmotility that characterizes diabetes. [ABSTRACT FROM AUTHOR]

Background: Poekilocerus pictus (Fabricius 1771), a painted grasshopper, sequesters cardenolides from its food plant, the Apple of Sodom or Aak, Calotropis procera (Aiton) W.T. Aiton (Family-Asclepiadaceae). In our present investigation, we were able to isolate Pseudomonas aeruginosa KRK6 from the intestine of Poekilocerus pictus responsible for the biotransformation of cardenolides. Methods: Pseudomonas aeruginosa KRK6 was grown in methanolic extracts of Calotropis procera and the modified cardenolides were detected by Liquid Chromatography-Mass Spectrometry (LCMS) and also used to induce apoptosis in cancer cells (MCF-7 cells and T-47 D). Result: The modified cardenolides CPMEP6 was found to induce apoptosis in human breast adenocarcinoma cells (MCF cells-IC50= 6.31±0.4 µg/mL, T-47D cells-IC50= 10.1±1.02 µg/mL). Phosphatidylserine exposure and DNA fragmentation suggested apoptosis in treated cancer cells. CPMEP6 induced apoptosis in cancer cells via the mitochondrial pathway by down-regulating BCL-2 protein expression and up-regulating BAX protein expression. [ABSTRACT FROM AUTHOR]

In the present study, we utilized db/db mice to investigate the mechanisms of apoptosis in brain and retinal neurons. A total of 30 male db/db mice aged 8–9 weeks were randomly assigned to the model group, Dihuangyinzi decoction (DHYZ) group (30.03 g/kg), and metformin (MET) group (0.58 g/kg), with 10 mice in each group. The control group comprised 10 db/m mice of the same background and age. Paired-associate learning (PAL) tests were conducted to evaluate the learning and memory functions of the mice. Histological assessments, including Hematoxylin-Eosin (H&E) and Nissl staining, were employed to observe changes in nerve cells in the hippocampus and retina. Immunohistochemistry and real-time fluorescence quantitative PCR were employed to detect the positive expression of anti-apoptotic factor Bcl-2 and pro-apoptotic factor Bax at the protein and mRNA levels in the hippocampus and retina. Western blotting analysis was adopted to assess protein expression levels of JNK, p-JNK, p38 MAPK, and p-p38 MAPK. Results revealed a significant decline in the correct rate of PAL test results in the model group (P < 0.001), accompanied by increased reaction and delay times (P < 0.001) and higher blood glucose levels (P < 0.001). H&E and Nissl staining indicated a reduction in the number of nerve cells in the CA1 area of the hippocampus in the model group, with scattered arrangement and decreased Nissl corpuscles. Positive expression and mRNA expression of Bax in the hippocampus and retina increased significantly (P < 0.001), while positive expression and mRNA expression of Bcl-2 decreased (P < 0.001). Protein expression levels of p-JNK/JNK and p-p38 MAPK/p38 MAPK showed a significant increase (P < 0.001). The DHYZ and MET groups exhibited enhanced accuracy in PAL experiments (P < 0.05), decreased reaction time and delay time (P < 0.05), and reduced blood glucose levels (P < 0.05). The number of neurons in the CA1 region of the hippocampus increased, morphological structure improved, and the arrangement of the hippocampal structure became more orderly. Additionally, no obvious vacuolization was observed in the neuronal cell layer. The number of Nissl bodies increased, and the layers of the retina were closely arranged, with an increase in the number of Nissl bodies. Positive expressions of Bax in the hippocampus and retina, both at the protein and mRNA levels, decreased significantly (P < 0.001), while positive expressions of Bcl-2 and its mRNA increased (P < 0.01, P < 0.001). Furthermore, the protein expressions of p-JNK/JNK and p-p38 MAPK/p38 MAPK decreased significantly (P < 0.01). This study suggested that DHYZ decoction could inhibit the JNK/p38 MAPK signaling pathway, thereby increasing the anti-apoptotic factor Bcl-2 and reducing the expression of the pro-apoptotic factor Bax, consequently inhibiting apoptosis in the hippocampal CA1 region and retinal neurons of db/db mice. [ABSTRACT FROM AUTHOR]

The pathological and physiological process of spinal cord injury is complex, and there is currently no effective treatment method. Magnetic stimulation is an emerging electromagnetic therapy method in recent years, and studies have shown its potential to reduce cell apoptosis. This study used an improved Allen's method to replicate an incomplete spinal cord injury rat model, and repetitive magnetic stimulation (rMS) intervention was performed on the rats for 21 days. The research plan consists of two parts. The first part aims to observe the effects of rMS on motor function and neuronal cell apoptosis in rats. The Basso-Beattie-Bresnahan (BBB) score results indicate that rMS promotes the recovery of motor function in rats; H&E staining showed that rMS improved spinal cord structural damage and inflammatory infiltration; TUNEL and NeuN staining suggest that rMS can reduce cell apoptosis and promote neuronal cell survival. The second part aims to explore the mechanism of action of rMS. Immunofluorescence staining showed that after rMS intervention, the positive counts of PI3K and Akt increased, whereas the positive counts of caspase-3 decreased. Western blot showed that after rMS intervention, the expression of phospho-phosphatidylinositol-3 kinase (p-PI3K)/PI3K, phospho (p)-Akt/Akt, and Bcl-2 increased, whereas the expression of Bcl-2-associated X protein (Bax) and caspase-3 decreased. In summary, rMS can significantly reduce cell apoptosis in the damaged spinal cord and promote neuronal cell survival. Its mechanism of action may be related to promoting the expression of PI3K/Akt pathway proteins, upregulating the antiapoptotic protein Bcl-2, downregulating the proapoptotic protein Bax, and thereby inhibiting the expression of apoptotic protein caspase-3. NEW & NOTEWORTHY: Spinal cord injury is a serious disabling central nervous system disease. Recently, research on magnetic stimulation therapy for spinal cord injury has been increasing, and its potential has gradually attracted the attention of experts. This study found that repetitive magnetic stimulation (rMS) can improve motor function and reduce neuronal apoptosis in spinal cord injury rats. The mechanism may be related to increasing the expression of phosphatidylinositol-3 kinase (PI3K)/Akt protein, thereby inhibiting cell apoptosis and promoting neuronal survival. [ABSTRACT FROM AUTHOR]