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Introduction: Migraine, a common disorder marked by severe and repetitive headaches, has been linked to the involvement of the NMDA receptor (NMDAR), a receptor responsible for glutamate signaling. Moringa oleifera (M. oleifera), recognized for its anti-inflammatory properties and therapeutic potential in various conditions, has been investigated. This study aims to assess the efficacy and precise mechanisms of M. oleifera for the treatment of migraine, for which evidence is limited. Methods: Rats were stratified into four distinct groups. The control group did not undergo the migraine-induction protocol. Post-induction, the "sumatriptan" group was administered sumatriptan injections, the "treatment" group received oral M. oleifera extract, and the "vehicle" group was provided with oral solvent treatment. Behavioral evaluations encompassing Von Frey's and hot plate assessments, in addition to qPCR analysis targeting Nr2a, Nr2b, Bax, Bcl-2, and Caspase-3, were conducted. Results: Von Fery's and hot plate tests revealed a notable decrease in triggering pressure and temperature within the vehicle group when compared to the other groups (both ps < 0.001). The Nr2a expression levels in both the vehicle and treatment cohorts exhibited significantly higher values than those observed in the control group (p < 0.001, p = 0.001) and the sumatriptan group (p < 0.001, p = 0.002). Conversely, no substantial alterations in Nr2b or Bcl-2 expression levels were observed across the study groups (p = 0.404, p = 0.976). Notably, heightened expressions of Caspase-3 and Bax were evident in the vehicle group relative to the other groups (p = 0.013, p = 0.010). Conclusions: Moringa oleifera extract appears to mitigate symptoms of migraine by inhibiting apoptosis, suggesting potential efficacy in migraine treatment; however, additional research investigating a wider range of pathways is necessary. Clinical trial number: Not applicable. [ABSTRACT FROM AUTHOR]

Alzheimer's disease (AD) is the most prevalent neurodegenerative disorder, characterized by the accumulation of amyloid-beta (Aβ) plaques and neuroinflammation. This study investigates the protective effects of noradrenaline (NA) on human microglial cells exposed to lipopolysaccharides (LPS) and Aβ aggregates—major contributors to inflammation and cellular damage in AD. The reduced Aβ aggregation in the HMC3 human microglial cells co-treated with Aβ and NA was confirmed by thioflavin T (ThT) assay, fluorescent ThT staining, and immunocytochemistry (ICC). The significantly increased viability of HMC3 cells after 48 h of incubation with NA at 50 µM, 25 µM, and 10 µM, exposed to IC50 LPS and IC50 Aβ, was confirmed by XTT and LDH assays. Moreover, we found that NA treatment at 25 μM and 50 μM concentrations in HMC3 cells exposed to IC50 LPS or IC50 Aβ results in an increased proliferation of HMC3 cells, their return to normal morphology, decreased levels of DNA damage, reduced caspase-3 activity, decreased expression of pro-apoptotic DDIT3 and BAX, and increased expression of anti-apoptotic BCL-2 genes and proteins, leading to enhanced cell survival, when compared to that of the HMC3 cells treated only with IC50 LPS or IC50 Aβ. Furthermore, we showed that NA induces the degradation of both extracellular and intracellular Aβ deposits and downregulates hypoxia-inducible factor 1α (HIF-1α), which is linked to impaired Aβ clearance and AD progression. These findings indicate that NA holds promise as a therapeutic target to address microglial dysfunction and potentially slow the progression of AD. Its neuroprotective effects, particularly in reducing inflammation and regulating microglial activity, warrant further investigation into its broader role in mitigating neuroinflammation and preserving microglial function in AD. [ABSTRACT FROM AUTHOR]

Mitochondria-targeting agents, known as mitocans, are emerging as potent cancer therapeutics due to pronounced metabolic and apoptotic adaptations in the mitochondria of cancer cells. ONC212, an imipridone-family compound initially identified as a ClpP agonist, is currently under investigation as a potential mitocan with demonstrated preclinical efficacy against multiple malignancies. Despite this efficacy, the molecular mechanism underlying the cell death induced by ONC212 remains unclear. This study systematically investigates the mitochondrial involvement and signaling cascades associated with ONC212-induced cell death, utilizing HeLa and A549 cancer cells. Treated cancer cells exhibited characteristic apoptotic features, such as annexin-V positivity and caspase-3 activation; however, these occurred independently of typical mitochondrial events like membrane potential loss (ΔΨm) and cytochrome c release, as well as caspase-8 activation associated with the extrinsic pathway. Additionally, ONC212 treatment increased the expression of anti-apoptotic proteins Bcl-2 and Bcl-xL, which impeded apoptosis, as the overexpression of Bcl-2-GFP and Bcl-xL-GFP significantly reduced ONC212-mediated cell death. Furthermore, combining a sub-lethal dose of the Bcl-2/Bcl-xL inhibitor Navitoclax with ONC212 markedly augmented caspase-3 activation and cell death, still without any notable ΔΨm loss or cytochrome c release. Moreover, inhibition of caspase-9 activity unexpectedly augmented, rather than attenuated, caspase-3 activation and the subsequent cell death. Collectively, our research identifies ONC212 as an atypical mitochondrial-independent, yet Bcl-2/Bcl-xL-inhibitable, caspase-3-mediated apoptotic cell death inducer, highlighting its potential for combination therapies in tumors with defective mitochondrial apoptotic signaling. [ABSTRACT FROM AUTHOR]

Epilepsy is one of the most common and serious diseases of the brain, affecting more than 70 million people worldwide. Available anticonvulsants are able to suppress seizures in two thirds of patients, and in the remaining third of patients, epilepsy is considered drug-resistant and other types of treatment are required, such as surgery, which also does not always lead to positive results. Overcoming resistance is a complex task that requires an understanding of the biochemical pathways and general pathological processes underlying epilepsy, primarily apoptosis. The purpose of this work was to study the effect of the antibiotic minolexin on the levels of apoptosis and the expression of apoptosis-associated molecules (p53, Bcl-2, caspase-3 and caspase-8) in the temporal cortex, underlying white matter and hippocampus of Krushinsky–Molodkina rats with hereditary audiogenic epilepsy with long-term kindling. We used Krushinsky–Molodkina rats at the age of 11 months, which were subjected to audiogenic stimulation and administered intraperitoneally with 1 mL of saline solution or the second-generation tetracycline series minolexin at a dose of 45 mg/kg, dissolved in saline solution for 14 days. The temporal lobe cortex and underlying white matter, the hippocampus, were examined. Apoptosis levels (TUNEL) and expression of apoptosis-associated proteins (p53, Bcl-2, caspase-3 and -8) were assessed (immunohistochemistry, Western blotting). In Krushinsky–Molodkina rats with hereditary audiogenic epilepsy, an increase in the apoptosis level was shown during long-term kindling. A p53-mediated, but caspase-independent mechanism of apoptosis activation has been identified. When minolexin was administered, an anti-apoptotic and neuroprotective effect was observed in the temporal lobe and hippocampus of rats. [ABSTRACT FROM AUTHOR]

Objectives: The purpose of this study was to determine the correlation between microscopic degeneration in the long head of the biceps tendon (LHBT) and the apoptotic process. Methods: This study included 26 consecutive patients who had undergone arthroscopic biceps tenodesis or tenotomy for symptomatic LHBT with or without concomitant rotator cuff tears (RCTs). Histological examination of the specimens under a light microscope was conducted after staining with hematoxylin, eosin, and the Alcian blue. Histopathological changes were assessed using the original Bonar score and the modified Bonar score and then correlated with the expression of the subsequent apoptosis markers: activated caspase-3 (casp3), tumor protein p53 (p53), and B-cell lymphoma 2 (BCL-2). Results: The mean original Bonar score was 8.65 (range 5–11), while the modified Bonar score was 7.61. There was no correlation between the original Bonar score and the age of the patients, but a positive correlation was found between the modified Bonar score and the age of the patients (p = 0.0022). There was no correlation between the age of patients and the expression indexes of BCL-2 and casp3. However, the expression of the p53 index showed a positive correlation with patient aging (p = 0.0441). Furthermore, there was no correlation observed between the expression of apoptotic indexes and both the original and modified Bonar scale. Conclusions: In LHB tendinopathy, the expression of apoptosis does not seem to directly correlate with the extent of degeneration, particularly in the late stages of tendinopathy. However, the transformations observed in collagen and ground substance were significantly associated with age, as well as tendinous tissue degeneration quantified according to modified Bonar score. The age of patients was also linked with the expression of the p53 index, as an increased apoptosis in the studied population. [ABSTRACT FROM AUTHOR]

Objective: There are numerous diagnostic, biological, and histological manifestations of salivary gland tumors, each of which offers concerns and difficulties in terms of diagnosis, grading, categorization, and therapy. The purpose of this study was to evaluate and compare the immunohistochemical expression of Bcl-2, caspase-3, and GSTP in benign and malignant salivary gland tumors, as well as how they are connected to a variety of clinicopathological variables. Methods: A total of 61 cases of buffered formalin-fixed, paraffin-embedded tissues from previously identified cases of benign and malignant salivary gland tumors were included in this study. The immunohistochemistry staining process was carried out according to the manufacturer's recommendations, employing polyclonal anti-Bcl-2, anti-caspase-3, and anti- GST antibodies. Results: The correlation between mean tumor diameter and Bcl-2 expression was shown to be statistically significant (rs=0.258, p<0.05). In pleomorphic adenoma tumor tissues, there were statistically significant correlations between the expression levels of Bcl-2 and caspase-3 (rs=0.66, p<0.01), Bcl-2 and GST (rs=0.61, p<0.01), and caspase-3 and GST (rs=0.73, p<0.01) when tumor types were compared. The tissues with pleomorphic adenoma, adenoid cystic carcinoma, and mucoepidermoid carcinoma had the highest staining intensity of Bcl-2 expression, while the lowest staining intensity of GSTP expression was observed. Conclusion: It seems probable to draw the conclusion that salivary gland tumors that resist apoptosis have elevated levels of Bcl-2 expression. The prognosis for salivary gland tumors may be poor due to the positive correlation between tumor diameter and high Bcl-2 expression. [ABSTRACT FROM AUTHOR]

One of the most common cancers in the world is oral squamous cell carcinoma. Increasing apoptotic activity is one method of treating cancer by reducing or stopping the uncontrolled growth of cancer cells. Known for its many health advantages and anti-tumor capabilities, Ant's Nest (Myrmecodia pendans) is a native of Papua, Indonesia. This study aimed to analyze the therapeutic effects of Ant's Nest extract on epithelial dysplasia in a carcinoma model through the immunoexpression of Caspase-3, Bax, and Bcl-2. The research was a pure experimental study using male Sprague Dawley rats, divided into control and treatment groups. The experimental animals were induced with 0.5% dimethylbenz(a)anthracene (DMBA) for 4 weeks, resulting in epithelial dysplasia. Subsequently, Ant's Nest water and ethanol extracts were administered orally at a dose of 6.3 mg dissolved in 0.5% CMC once a day for 4 weeks. Immunohistochemical examinations were conducted for Caspase-3, Bax, and Bcl-2 immunoexpression. The results of the study showed that there were notable variations in the immunoexpression of Bcl-2, Bax, and Caspase-3 between the treatment and control groups. The group that was given Ant's Nest water and ethanol extracts showed a decrease in Bcl-2 immunoexpression and an increase in Caspase-3 and Bax immunoexpression. We find that via modulating the immunoexpression of Caspase-3, Bax, and Bcl-2, Ant's Nest extract treatment promotes the process of apoptosis. [ABSTRACT FROM AUTHOR]

Objective: To study the in vitro and in vivo antitumor effects of the polysaccharide of Alocasia cucullata (PAC) and the underlying mechanism. Methods: B16F10 and 4T1 cells were cultured with PAC of 40 µg/mL, and PAC was withdrawn after 40 days of administration. The cell viability was detected by cell counting kit-8. The expression of Bcl-2 and Caspase-3 proteins were detected by Western blot and the expressions of ERK1/2 mRNA were detected by quantitative real-time polymerase chain reaction (qRT-PCR). A mouse melanoma model was established to study the effect of PAC during long-time administration. Mice were divided into 3 treatment groups: control group treated with saline water, positive control group (LNT group) treated with lentinan at 100 mg/(kg·d), and PAC group treated with PAC at 120 mg/(kg·d). The pathological changes of tumor tissues were observed by hematoxylin-eosin staining. The apoptosis of tumor tissues was detected by TUNEL staining. Bcl-2 and Caspase-3 protein expressions were detected by immunohistochemistry, and the expressions of ERK1/2, JNK1 and p38 mRNA were detected by qRT-PCR. Results: In vitro, no strong inhibitory effects of PAC were found in various tumor cells after 48 or 72 h of administration. Interestingly however, after 40 days of cultivation under PAC, an inhibitory effect on B16F10 cells was found. Correspondingly, the long-time administration of PAC led to downregulation of Bcl-2 protein (P<0.05), up-regulation of Caspase-3 protein (P<0.05) and ERK1 mRNA (P<0.05) in B16F10 cells. The above results were verified by in vivo experiments. In addition, viability of B16F10 cells under long-time administration culture in vitro decreased after drug withdrawal, and similar results were also observed in 4T1 cells. Conclusions: Long-time administration of PAC can significantly inhibit viability and promote apoptosis of tumor cells, and had obvious antitumor effect in tumor-bearing mice. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To study the effect of dictyophora rubrovalvata polysaccharide on proliferation and apoptosis of human gastric cancer MKN-45 cells, and probe into the possible mechanism based on mitogen activated protein kinase (MAPK) signaling pathway. METHODS: MKN-45 cells were cultured in vitro, and divided into the control group, low-dose, medium-dose and high-dose dictyophora rubrovalvata polysaccharide group. The effect of dictyophora rubrovalvata polysaccharide with final concentrations of 0 mg / mL, 4 mg / mL, 6 mg / mL and 8 mg / mL on the proliferation of human gastric cancer MKN-45 cells was detected by cell counting (CCK-8) after 48 h of treatment. The morphological changes of MKN-45 cells were observed under inverted microscope after being treated with different concentrations of dictyophora rubrovalvata polysaccharide for 48 h. In the experimental group, the final concentration of polysaccharide was 8 mg / mL. Apoptosis rate was detected by Annexin V-FITC / PI double staining flow cytometry. Cell cycle was detected by flow cytometry. Western blotting was used to detect the expression of p38 protein kinase (p38), phosphorylated p38 protein kinase ( p-p38), extracellular regulated protein kinase (ERK), phosphorylated extracellular regulated protein kinase (p-ERK), C-JUN N-terminal kinase (JNK), phosphorylated C-JUN N-terminal kinase (p-JNK), B-lymphomatoma-2 protein (Bcl-2) and the activated cysteine proteinase-3 ( cleaved Caspase-3) protein, by the ratio of gray value to glyceraldehyde 3-phosphate dehydrogenase (GADPH). RESULTS: After 48 h of treatment, the inhibition rates of dictyophora rubrovalvata polysaccharide under different concentration ( 4 mg / mL, 6 mg / mL and 8 mg / mL) on the proliferation of human gastric cancer MKN-45 cells were (8. 27±5. 99)%, (21. 83± 5. 77)% and (56. 95±5. 21)%, respectively. In the detected concentration range, the inhibition rates of dictyophora rubrovalvata polysaccharide on MKN-45 cell proliferation increased with the increase of concentration. Under the inverted microscope, the MKN-45 cell density decreased significantly compared with the control group after being treated with 8 mg / mL dictyophora rubrovalvata polysaccharide for 48 h. Results of flow cytometry showed that compared with the control group, the apoptosis rate of MKN-45 cells in the experimental group increased significantly (P<0. 05); results of cell cycle showed that the proportion of cells in G1 phase and S phase in the experimental group decreased significantly ( P < 0. 05, P < 0. 05 ), and the proportion of cells in G2 phase increased significantly (P<0. 05); Western blotting results showed that compared with the control group, the cleaved Caspase-3 in MKN-45 cells in the experimental group increased ( P < 0. 05), Bcl-2 tended to decreased ( P < 0. 05), p-P38 / P38 ratio, p-ERK/ ERK ratio and p-JNK/ JNK ratio in the control group were lower than those in the control group (P<0. 05), with statistically significant differences. CONCLUSIONS: Dictyophora rubrovalvata polysaccharide can inhibit the proliferation and induce apoptosis of human gastric cancer MKN-45 cells in vitro, and block tumor cells in G2 / M phase, which may be achieved by regulating MAPK signaling pathway. [ABSTRACT FROM AUTHOR]

Brain insulin resistance is linked to metabolic syndrome (MetS). A low-carbohydrate, high-fat (LCHF) diet has been proposed to have a protective effect. Therefore, this study aimed to investigate the brain insulin resistance markers in a rat animal model of MetS and the protective effects of the LCHF diet. Four groups of male rats (10/group) were created. Group I (Control) was fed a regular diet. Groups II–IV were injected with dexamethasone (DEX) to induce MetS. Group II received DEX with a regular diet. Group III (DEX + LCHF) rates were fed a low-carbohydrate, high-fat diet, while Group IV (DEX + HCLF) rats were fed a high-carbohydrate, low-fat (HCLF) diet. At the end of the four-week experiment, HOMA-IR was calculated. Moreover, cerebral gene expression analysis of S-100B, BDNF, TNF-α, IGF-1, IGF-1 R, IGFBP-2, IGFBP-5, Bax, Bcl-2, and caspase-3 was carried out. In the DEX group, rats showed a significant increase in the HOMA-IR and a decrease in the gene expression of IGF-1, IGF-1 R, IGFBP-2, IGFBP-5, BDNF, and Bcl2, with a concomitant rise in S100B, TNF-α, Bax, and caspase-3. The LCHF diet group showed a significantly opposite effect on all parameters. In conclusion, MetS is associated with dysregulated cerebral gene expression of BDNF, S100B, and TNF-α and disturbed IGF-1 signaling, with increased apoptosis and neuroinflammation. Moreover, the LCHF diet showed a protective effect, as evidenced by preservation of the investigated biochemical and molecular parameters. [ABSTRACT FROM AUTHOR]

Research demonstrated that folate deficiency in either the mother or father could impact the biological functions of the offspring's of neural cells. Folate deficiency can also impair the methionine cycle, thus contributing to the conversion of S-adenosylmethionine (SAM) to S-adenosylhomocysteine (SAH), which could potentially cause damage to the central nervous system. The study focused on the effect of parental folate deficiency on neural cell apoptosis in offspring neonatal rats and whether it is mediated by the levels of SAM and SAH in brains. The experimental design was conducted by feeding female and male Sprague Dawley (SD) rats with either folate-deficient or folate-normal diets, sacrificing the offspring within 24 h and isolating their brain tissue. Rats were divided into four groups: the maternal-folate-deficient and paternal-folate-deficient (D-D) group; the maternal-folate-deficient and paternal-folate-normal (D-N) group; the maternal-folate-normal and paternal-folate-deficient (N-D) group; and the maternal-folate-normal and paternal-folate-normal (N-N) group. There was down-regulation of B-cell lymphoma 2 (Bcl-2) expression, up-regulation of Bcl-2-associated X protein (Bax) and Caspase-3 expression of neural cells, and pathological changes in the brain ultrastructure, as well as decreased SAM levels, increased SAH levels, and a decreased SAM/SAH ratio in the rat fetal brain via parental folate deficiency. In conclusion, parental folate deficiency could induce the apoptosis of neural cells in neonatal offspring rats, while biparental folate deficiency had the greatest effect on offspring, and the unilateral effect was greater in mothers than in fathers. This process may be mediated by the levels of SAM and SAH in the rat fetal brain. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the regulatory role of Aldo-keto reductase family 1 member B1 (AKR1B1) in glioma cell proliferation through p38 MAPK activation to control Bcl-2/BAX/caspase-3 apoptosis signaling. AKR1B1 expression was quantified in normal human astrocytes, glioblastoma multiforme (GBM) cell lines, and normal tissues by using quantitative real-time polymerase chain reaction. The effects of AKR1B1 overexpression or knockdown and those of AKR1B1-induced p38 MAPK phosphorylation and a p38 MAPK inhibitor (SB203580) on glioma cell proliferation were determined using an MTT assay and Western blot, respectively. Furthermore, the AKR1B1 effect on BAX and Bcl-2 expression was examined in real-time by Western blot. A luminescence detection reagent was also utilized to identify the effect of AKR1B1 on caspase-3/7 activity. The early and late stages of AKR1B1-induced apoptosis were assessed by performing Annexin V-FITC/PI double-staining assays. AKR1B1 expression was significantly downregulated in glioma tissues and GBM cell lines (T98G and 8401). Glioma cell proliferation was inhibited by AKR1B1 overexpression but was slightly increased by AKR1B1 knockdown. Additionally, AKR1B1-induced p38 MAPK phosphorylation and SB203580 reversed AKR1B1′s inhibitory effect on glioma cell proliferation. AKR1B1 overexpression also inhibited Bcl-2 expression but increased BAX expression, whereas treatment with SB203580 reversed this phenomenon. Furthermore, AKR1B1 induced caspase-3/7 activity. The induction of early and late apoptosis by AKR1B1 was confirmed using an Annexin V-FITC/PI double-staining assay. In conclusion, AKR1B1 regulated glioma cell proliferation through the involvement of p38 MAPK-induced BAX/Bcl-2/caspase-3 apoptosis signaling. Therefore, AKR1B1 may serve as a new therapeutic target for glioma therapy development.

Background and Aim: Sertraline is an antidepressant drug and many controversial reports have been presented on the complications of sertraline on the reproductive system. In this study we evaluated the protective effects of vitamin E against sertraline-induced damage to the reproductive system of male mice. Materials and Methods: In this experimental study, 40 adult male mice were divided into 8 groups of 5. Four groups received 100 IU/kg.bw vitamin E by gavage for 42 days. One group was considered as control and three groups received just sertraline with doses of 5, 10 and 20 mg/kg.bw orally. 24 hours after the last treatment, testes tissue and blood samples were collected and sent for histochemical and biochemical studies, spermatogenic indices as well as measurement of the expressions of Bcl-2, Caspase-3 and Hsp70-2 genes. Results: The results showed a significant increase in the level of malondialdehyde, nitric oxide and expression of Caspase-3 and Hsp70-2 genes. We also found a significant decrease in spermatogenesis indices, total antioxidant capacity, testosterone and Bcl-2 gene expression in the group receiving 20 mg/kg.bw sertraline (P

Thymoquinone (TQ), a polyphenol obtained from Nigella sativa plays a fundamental role in controlling homeostasis. It possesses anti-cancer, anti-oxidant, anti-inflammatory, and analgesic properties. Its anti-cancer properties extend to lymphoma, leukaemia, medulloblastoma, prostate, pancreatic, breast, and ovarian cancer (OvCa). OvCa is one of the most lethal malignancies worldwide having distinct symptoms and a poor prognosis. Resistance to Cisplatin (CDDP) and subsequent reoccurrence of tumors lead to treatment failure. To tackle this, combination therapies along with polyphenols are being studied and have been proven to enhance CDDP response. Certain proteins such as Bcl-2, Bax, STAT3, p53, and caspase-3/9 are involved in cancer progression and signaling of chemoresistance. In this study, Thymoquinone (TQ) interactions with these proteins have been observed through molecular docking. The 3D structure prediction was carried out using Phyre2 and SWISS-MODEL. The molecular docking of protein targets and TQ was done using the PatchDock server. The docked results were then visualized using Discovery Studio. This study helps us to understand the interactions between TQ and apoptotic proteins that help in the augmentation of conventional therapy. This knowledge will also help in developing a new approach and alternate therapeutic strategies using polyphenols like TQ, even in resistant cells. [ABSTRACT FROM AUTHOR]

Background and Objective: There is a growing need to comprehend the potential outcomes of nanoparticles (NPs) on human well-being, including their potential for detecting and treating leukemia. This study examined the role of iron folate core–shell and iron oxide nanoparticles in inducing apoptosis and altering the expression of the B-cell lymphoma 2 (Bcl-2), Bcl-2 associated X-protein (Bax), and Caspase-3 genes in leukemia cells. Methods: The obtained iron oxide and iron folate core–shell nanoparticles were analyzed using a variety of analytical techniques, including ultraviolet-visible (UV-Vis) absorption spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), dynamic light scattering (DLS), zeta potential, and transmission electron microscopy (TEM). Additionally, FTIR and UV-Vis were used to characterize doxorubicin. The MTT test was utilized to investigate the cytotoxicity of iron oxide and iron folate core–shell nanoparticles. The expression of the apoptotic signaling proteins Bcl-2, Bax, and Caspase-3 was evaluated using the real-time reverse transcription polymerase chain reaction (RT-qPCR) method. Additionally, flow cytometry was performed to gauge the degrees of necrosis and apoptosis. Results: UV-Visible spectroscopy analysis showed that the generated iron oxide and iron folate core–shell NPs had a distinctive absorption curve in the 250–300 nm wavelength range. The XRD peaks were also discovered to index the spherical form with a size of less than 50 nm, which validated the crystal structure. The FTIR analysis determined the bonds and functional groups at wavenumbers between 400 and 4000 cm-1. A viable leukemia treatment approach is a nanocomposite consisting of iron and an iron folate core-shell necessary for inhibiting and activating cancer cell death. The nearly resistant apoptosis in the CCRF-CEM cells may have resulted from upregulating Bax and Casepase-3 while downregulating Bcl-2 expression. Conclusions: Our study documents the successful synthetization and characterization of iron oxide, which has excellent anticancer activities. A metal oxide conjugation with the nanoparticles’ core–shell enhanced the effect against acute leukemia.

Apoptosis, programmed cell death, has a central role in developmental biology and in maintaining the equilibrium of renewing tissues. A founding member of the Bcl-2 family of regulatory proteins for apoptosis is Bcl-2, which is encoded by the BCL2 gene. Caspase-3 shares typical features with all caspases, including the role of acting as a crucial mediator of apoptosis.

Introduction: In the present study, we aimed to test the hypothesis that hypercapnia, independently and/or in combination with hypoxia, can activate signaling pathways related to the inhibition of proapoptotic (caspase-dependent and caspase-independent) factors and the induction of antiapoptotic factors in facilitating adaptation to hypoxia/ischemia. Materials and methods: Following exposure to permissive hypercapnia and/or normobaric hypoxia, the degree of apoptosis was evaluated in experimental ischemia models in vivo and in vitro. The percentages of caspase-3, apoptosis-inducing factor (AIF), Bax, and Bcl-2 in astrocytes and neurons derived from male Wistar rats were also calculated. In vitro, cells were subjected to various types of respiratory exposure (hypoxia and/or hypercapnia for 24 or 12 h) as well as further sublethal chemical hypoxia. The percentages of these molecules in nerve cells in the ischemic penumbra of the brain after photothrombotic injury were also calculated. Results: The degree of apoptosis was found to decrease in ischemic penumbra, mostly due to the hypercapnic component. It was also discovered that the levels of caspase-3, AIF, and Bax decreased in this region, whereas the Bcl-2 levels increased following exposure to hypercapnia and hypercapnic hypoxia. Conclusions: This integrative assessment of the rate of apoptosis/necrosis in astrocyte and neuron cultures shows that the combination of hypercapnia and hypoxia resulted in the maximum neuroprotective effect. The levels of apoptosis mediators in astrocyte and neuron cultures were calculated after modeling chemical hypoxia in vitro. These results show that the exposure models where permissive hypercapnia and normobaric hypoxia were combined also had the most pronounced inhibitory effects on apoptotic signaling pathways. [ABSTRACT FROM AUTHOR]

Background and Aims: Amanita phalloides is the deadliest toxic mushroom in the world and causes death from acute liver failure. α-amanitin (α-AMA), the most potent toxin, inhibits RNA polymerase II in hepatocytes, stops protein synthesis, and causes hepatotoxicity. However, the information about the mechanisms underlying hepatotoxicity caused by α-AMA is quite inadequate. This study aims to reveal the complex necrotic and apoptotic mechanisms occurring in mouse hepatocytes depending on A. phalloides exposure time in vivo. Methods: BALB-c male mice were divided into 5 groups (n=7): control, α-AMA-2, α-AMA-12, α-AMA-72, and α-AMA-96 groups. A poisoning model was created by oral administration of A. phalloides mushroom extract containing 10 mg/kg of α-AMA to mice and they were sacrificed after 2, 12, 72, and 96 h. Then, TNF-α, Bax, caspase-3, and Bcl-2 gene expression levels in liver tissues were examined by the RT-qPCR method. Time-dependent damage to liver tissues was also evaluated histopathologically. Results: RT-qPCR results showed that proinflammatory cytokine TNF-α mRNA expression levels increased in mouse liver tissues at 2 and 12 h after A. phalloides administration compared among the groups. Bax mRNA expression levels increased in the 12 and 72 h after A. phalloides ingestion. It was observed that caspase-3 mRNA expression levels increased in the 72 and 96 h groups compared among the groups, while Bcl-2 mRNA expression levels decreased in the 72 and 96 h groups. Conclusion: Our findings showed that necrotic mechanisms develop in the early period after A. phalloides mushroom poisoning, and then apoptotic mechanisms are effective. In conclusion, understanding the mechanisms of A. phalloides-induced hepatotoxicity will provide important information for new treatment strategies to be developed. [ABSTRACT FROM AUTHOR]

Ibuprofen is a nonsteroidal anti-inflammatory drug that is commonly used for its analgesic, antipyretic and anti-inflammatory effects worldwide. However ibuprofen comes with serious unavoidable adverse effects on various organs when used for long duration or overdosed. Trichopus zeylanicus is a medicinal plant endemic to India owning various beneficial properties and is been used in treating various ailments. Therefore, the objective of this study was to evaluate the ameliorative effect of aqueous leaves' extract of Trichopus zeylanicus against ibuprofen-induced hepatic toxicity and enteropathy in rats. Overall in this study 30 male albino rats were used, which were divided into five groups (six in each group). Group-I was normal control, Group-II was ibuprofen (400 mg/kg/day) inebriated group, Group-III was silymarin (25 mg/kg/day) pretreated + ibuprofen (400 mg/kg/day), Group-IV was ALETZ (1000 mg/kg/day) pretreated + ibuprofen (400 mg/kg/day), and Group-V was ALETZ alone (1000 mg/kg/day) group. The duration of the administration was for five days, followed by scarifying rats on the sixth day. Later the rats were assessed for liver and intestine enzyme markers, antioxidant parameters along with histopathological changes. In addition the pro-inflammatory markers such as TNF-α, IL-6 and IL-1β as well as anti-inflammatory cytokine IL-10 levels were measured using ELISA. Lastly the expression pattern of apoptotic signaling markers such as caspase-3, caspase-8 and Bcl-2 was evaluated using western blot. The results obtained from this study showed changes in levels of aforesaid parameter which presented the toxic effect of ibuprofen on liver and small intestine. Pre-treatment of ALETZ in ibuprofen-inebriated group was able to normalize the adverse effect caused due to ibuprofen. The conclusion of the study deduces that pre-treatment with ALETZ alleviates by modulating oxidative stress, inflammation, and apoptosis in ibuprofen inebriated rats, indicating its protective mechanism. [ABSTRACT FROM AUTHOR]

Definition: Apoptosis, programmed cell death, has a central role in developmental biology and in maintaining the equilibrium of renewing tissues. A founding member of the Bcl-2 family of regulatory proteins for apoptosis is Bcl-2, which is encoded by the BCL2 gene. Caspase-3 shares typical features with all caspases, including the role of acting as a crucial mediator of apoptosis. [ABSTRACT FROM AUTHOR]

Objective: The aim is to investigate the protective effect of andrographolide (AP) on retinal ganglion cells (RGC) in rats with glaucoma and its mechanism. Methods: ninety-six adult male Wistar rats were randomly divided into normal control (NC) group, chronic ocular hypertension (CHOT) group, vehicle group, and AP group. The density of RGCs and the number of apoptotic cells in retinal slices were evaluated, and the function of RGC was evaluated by photopic negative wave response (PhNR). The expression of Bcl-2, Bax, and caspase-3 in the retina was detected. Results: (1) The density of RGC in the CHOT and vehicle groups were significantly lower than that in the NC and AP (P < 0.01). After AP intervention, the RGC density of the rats significantly increased (P < 0.01). The ganglion cell layer (GCL) of the CHOT and vehicle groups was obviously thinner, and the thickness of the GCL was partially restored in the AP group. (2) A large number of terminal deoxytransferase mediated dUTP nick end labeling (TUNEL) positive cells were found in the GCL of the CHOT and vehicle groups, but only a few TUNEL positive cells were found in the AP group. The percentage of TUNEL positive cells was 8.80 ± 4.97%, 37.00 ± 5.27%, 46.16 ± 6.50%, and 22.29 ± 3.52% for groups NC, CHOT, vehicle, and AP, respectively. (3) At 2 weeks, the amplitude of PhNR in the CHOT and vehicle groups decreased by 63.22 ± 13.89% and 57.88 ± 6.95%, respectively. The amplitude of PhNR in the AP group was only 22.56 ± 6.44% lower than that in the NC group. The AP intervention significantly reduced the decrease of the PhNR amplitude in CHOT eyes. (4) Compared with the NC group, the expression of anti-apoptotic protein Bcl-2 was decreased in the CHOT and vehicle groups, and the expression of the pro-apoptotic protein Bax and caspase-3 significantly increased in the CHOT and vehicle groups, which could be reversed by AP treatment. The trend of the reverse transcription-polymerase chain reaction was consistent with that of the western blot analysis. Conclusion: The protective effect of caspase-3/Bcl-2 may be achieved through the regulation of the Caspase-3/Bcl-2 pathway in the rat retina.

Is there a relationship between embryo quality, pregnancy rates and apoptotic gene expression in cumulus cells of oocytes collected from patients with poor ovarian response and polycystic ovary syndrome? Fifty infertile couples who underwent assisted reproductive technology treatment were included in the study (Approval date 4 February 2020, number 03). The patients were divided into four group: control (n = 9; 90 oocytes), unexplained infertility (n = 8; 86 oocytes), polycystic ovary syndrome (PCOS) (n = 6; 137 oocytes) and poor ovarian response (POR) (n = 27; 124 oocytes). Cumulus cells were isolated individually from 437 oocytes obtained. Intracytoplasmic sperm injection was undertaken on 365 mature oocytes. The embryos were monitored. Caspase-3, Bax and Bcl-2 gene expressions of the cumulus cells were measured by real-time polymerase chain reaction. A significant and negative correlation was found between Bax and Bcl-2 expressions of the cumulus cells of poor-quality embryos. The increase in Caspase-3 gene expression in the POR group statistically decreases the pregnancy rates. Fertilization and good-quality embryo development of 365 oocytes whose cumulus cells were examined, however, were not associated with apoptotic gene expression. The Bax/Bcl-2 ratio was found to be significantly lower in cumulus cells of mature oocytes. Our results demonstrated no significant associations between fertilization, quality embryo development and apoptotic gene expression. Bax expression and the Bax/Bcl-2 ratio are high in immature oocyte cumulus cells has shown us that the apoptotic process may begin when the cumulus–oocyte connection exists. [ABSTRACT FROM AUTHOR]

Objective electroencephalogram ( EEG) discharge index in patients with traumatic epilepsy and EEG abnormalities. Methods68 patients with traumatic epilepsy treated in Haian People’s Hospital from January 2019 to January 2021 were selected. According to the EEG test results, there were 11 with normal EEG ( EEG normal group), 20 with EEG boundary linearity ( boundary linearity group), 21 with abnormal EEG slow wave ( slow wave abnormal group) and 16with epileptic discharge ( epileptic discharge group) . The relative expression of plasma microrna-7 ( miR-7) was calculated by 2-ΔΔCt, and the contents of apoptotic molecules Bcl-2, Bax and caspase-3 were measured by enzymelinked immunosorbent assay ( ELISA) . EEG was examined by electroencephalograph, and EEG discharge index was calculated. The data correlation of normal distribution was analyzed by Pearson correlation analysis and multiple linear regression analysis. Results There was a statistically significant difference in the levels of miR-7, Bcl-2, Bax, and caspase-3 among the groups ( P ＜ 0. 05), and with the increase of EEG abnormalities, the levels of miR-7 and Bcl-2decreased, the levels of Bax and caspase-3 increased, and the EEG firing index increased. The results of Pearson correlation analysis showed that there were correlations between miR-7, Bcl-2, Bax, and caspase-3 levels and EEG firing index in patients with traumatic epilepsy ( P ＜ 0. 05) . The results of multiple linear regression analysis showed that there were correlations between miR-7, Bcl-2, Bax, and caspase-3 levels and EEG abnormalities ( P ＜ 0. 05) .Conclusions MiR-7 and apoptotic molecules are correlated with EEG discharge index and EEG abnormalities in patients with traumatic epilepsy. Early EEG examination should be performed. The prognosis should be judged. A scientific and reasonable treatment plan should be made in combination with the patient’s medical history, course of disease and the results of miR-7 and apoptotic molecule detection. [ABSTRACT FROM AUTHOR]

Synthetic pyrethroids are a group of insecticides frequently used in public health and agriculture, and 3-PBA is a common metabolite of them. Although the liver is the primary organ responsible for metabolizing many compounds including pesticides, to the authors' knowledge there have been no studies on the direct hepatotoxic effects of 3-PBA. Therefore, this study aimed to investigate the possible hepatotoxic effects of 3-PBA on a Human Hepatoma Cell Line (HepG2) and the underlying apoptotic mechanisms. Firstly, an LC50 of 1041.242 µM was calculated for 3-PBA by using the WST-1 test with concentrations ranging between 1 µM and 10 mM. Following that, the HepG2 cells in the experimental group were exposed to 3 different concentrations of 3-PBA (1/5 LC50, 1/10 LC50 and 1/20 LC50) for 24 hours. The apoptotic mechanism was evaluated by using flow cytometry, and immunofluorescence assays for Caspase 3 and Bcl-2. In the flow cytometry assay, the total number of apoptotic cells increased in a dose dependent manner (p < 0.05). In the immunofluorescence assay, the Caspase 3 protein showed strong immunoreactivity in the experimental groups, while the reaction to the Bcl-2 protein was minimal. These results demonstrated that 3-PBA has a significant hepatotoxic effect on HepG2 cells and induces apoptosis via the regulation of Caspase-3 and Bcl-2. Furthermore, our results could further the understanding of the fundamental molecular mechanisms of 3-PBA hepatotoxicity. More studies are needed to determine the effects of long-term exposure to 3-PBA and also the molecular mechanisms underlying hepatotoxicity. [ABSTRACT FROM AUTHOR]

Background and Objectives: Aspartame is a non-nutritive and artificial sweetener which is widely used in diet and low calorie products and also in a variety of foods, drugs and hygiene products. The present study has been conducted in order to evaluation of the effects of aspartame on ovaries in female mice. Methods: In this experimental study, 36 adult female mice were randomly divided into four groups of nine each. For three groups aspartame was administered orally with the doses of 40, 80 and 160 mg/kg.BW respectively, and one group as the control group received normal saline for 91 days by gavage. Also a control group was considered. 24 hours after the last treatment, ovarian tissue and blood samples were collected and used for biochemical, histomorphological, histomorphometric, histochemical and gene expression studies. The obtained data were analyzed using One-Way Analysis of Variance (ANOVA) and Tukey’s test at the significance level of P

Objectives: The main objective of the present research is to provide GC–MS and LC-MS phytochemcial profiling of Corchorus olitorius along with Flow cytometry based mechanistic investigation of cytotoxic activity of its extracts against breast and lung cancer cell lines. Methods: The Soxhlet extraction method was used in the sequential extraction of C. olitorius. The phytochemical profiling was done using LC-MS and GC–MS techniques, and total phenolic and flavonoid content was determined. The cytotoxicity was studied against non-cancerous L929 cells and cancerous MCF-7 and A549 cell lines using MTT assay, followed by flow cytometry based molecular mechanisms study through anti-Bcl-2 assay, Caspase-3, and DNA fragmentation (TUNEL assay). Results: The C. olitorius methanolic extract showed the phenolic compounds and flavonoids as a major constituents with 699 µg GAE/g of dw and 1361.50 µg QE/g of dw, respectively. The GC–MS and LC-MS results confirmed the presence of active phytochemicals. Toxicity study against L929 cell line revealed that methanol extract proved to be less toxic with a higher IC50 value i.e. 227.84 µg/ml. Cell viability MTT assay for methanol extract against MCF-7 and A549 cell lines revealed significant results with IC50 values of 20 µg/ml and 12.45 µg/ml, respectively. The flow cytometry based molecular marker studies showed that the extract successively induced early and late apoptosis in tested breast and lung cancer cells through activation of Caspase-3 with inhibition of Bcl-2 protein and induced cell death through DNA damage. Conclusion: Collectively, these findings show that the methanol extract of C. olitorius inhibits breast cancer and lung cancer cell lines with significant cytotoxic activity. Thus, C. olitorius would be a promising source of chemopreventive agents that warrant further investigation to find lead compounds with cancer chemotherapeutic potential.

Boswellia sacra oleo gum resin (Burseraceae) commonly known as frankincense is traditionally used in many countries for its beneficial effect on male fertility. This study explores its effect on the male reproductive system after a 60-day repeated administration at two different doses to rats (in vivo) and on human Leydig cells (in vitro). The methanolic extract of B. sacra was analyzed for the presence of various constituents by preliminary phytochemical analysis and gas chromatography-mass spectrometry (GC-MS) while quantitative analysis of boswellic acids was done by high-performance liquid chromatography (HPLC). Administration of B. sacra extract to rats elevated the serum testosterone levels with an associated reduction in serum levels of FSH and LH. An increase in the activity of antioxidant enzymes, superoxide dismutase and catalase, was seen. A dose-dependent increase in the sperm count and sperm motility was also observed. The in vivo results were supported by changes in the expression of the Bcl-2 gene and caspase-3 gene in human Leydig cells in vitro. The results of this study support the traditional use of B. sacra to increase male fertility. [ABSTRACT FROM AUTHOR]

Objective To investigate the effect of esmolol, a short-acting, highly selective β-blocker, on cardiomyocyte apoptosis in septic rats and its possible mechanism. Methods A total of 63 male Wistar rats were randomly divided into sham operation group (Sham group), sepsis group (CLP group), and esmolol group (ES group). Each group was further divided into 6-, 12-, and 24-hour subgroups. The rats of the Sham group were given sham surgery only, and those of the CLP and ES groups underwent cecal ligation and puncture (CLP) to establish sepsis model. The Sham group and the CLP group were continuously infused with 0.9% sodium chloride solution at a speed of 1 mL/h by micropump for 6 h, while the ES group was continuously infused with 15 mg/kg·h esmolol diluted solution at 1 mL/h for 6 h. Then the rats were sacrificed at 6, 12 and 24 h after successful modeling, and the heart specimens were collected. Serum TNF-α and BNP levels were measured by ELISA. Water soluble tetrazolium (WST-1) method was used to detect SOD activity, and spectrophotometry to determine MDA content in myocardial tissue. The expression levels of Caspase-3, Bcl-2, Bax and other proteins in myocardial tissue were detected by Western blotting. Transmission electron microscopy was performed to observe the morphology and structure of myocardial mitochondria. Results Compared with the levels at the same time points in Sham group, the serum TNF-α and BNP levels, MDA content, and expression levels of apoptotic proteins such as Caspase-3 and Bax were significantly increased in the CLP and ES groups (P < 0.05). Except the level of BNP in ES24h group was not significantly different than that in CLP24h group (P>0.05), ES group had obviously decreased serum TNF-α and BNP levels, MDA content, and expression levels of Caspase-3 and Bax when compared with those in the CLP group at same time points (P < 0.05). The SOD activity and Bcl-2 level were remarkably higher in the CLP and ES groups at the same time points (P < 0.05), and the activity and levels were increased in the ES group than the CLP group at all time points (P < 0.05). Under a transmission electron microscope, myocardial fibers and mitochondria were almost intact in the Sham group at all time points; the damages were the most severe in the CLP group, and the severity was deteriorated gradually with time elapse, while the conditions became alleviated in the ES group at each time point. Conclusion Esmolol partially improves cardiac function in septic rats by inhibiting cardiomyocyte apoptosis. The mechanism may be due to its inhibition in apoptosis initiation factor TNF-α and regulation in the expression of apoptosis-related proteins such as Bcl-2, Bax, and Caspase-3, and reduction of the oxidative stress response as well. It mediates both the accompanying process of apoptosis and an important initiator of apoptosis, and protects the mitochondrial structure of target organelles damaged by apoptosis and oxidative stress.

The goal of this investigation is to report the fabrication, characterization, cytotoxicity, and apoptotic assessment of new platinum based compounds on K562 and HL-60 human leukemia cells. Two new platinum (II) compounds, Pt-5a and Pt-6a, were prepared and characterized by fourier transform infrared spectroscopy (FTIR), proton nuclear magnetic resonance spectroscopy (1HNMR), environmental scanning electron microscopy (ESEM) and energy dispersive spectrometer (EDS) techniques. The cytotoxic activities of the compounds were evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) test. Caspase-3, B-cell lymphoma 2 (Bcl-2), and B-cell lymphoma 2 associated X protein (Bax) gene expressions were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) to illuminate the mechanism of apoptosis. The results present that the applied compounds exhibited dose-dependent cytotoxic effects in-vitro. Pt-5a and Pt-6a compounds caused a rise in Bax in HL-60 cells while a reduction in Bcl-2 was recorded in all applied doses. In HL-60 cells, an increase in caspase-3 was detected at doses of 25 µM and 50 µM of Pt-5a and 30 µM of Pt-6a. The treatment with 40 µM of Pt-5a increased caspase-3 and Bax in K562 cells compared with control cells. Bcl-2 was found to be low in 20 µM of Pt-5a treatment in K562 cells. Pt-6a caused a significant increase in caspase-3 at the dose of 30 µM in the same cells. It is proposed that the newly synthesized platinum compounds may prove to be significant in the development of anticancer-effective drugs as they trigger apoptosis in a dose-dependent manner. [ABSTRACT FROM AUTHOR]

Background: Doxorubicin (Dox) is one of the most effective antineoplastic drugs which has severe cardiotoxic effects, limiting its clinical usage. Though the exact mechanism of doxorubicin-induced cardiotoxicity is yet to be elucidated, it is shown that production of reactive oxygen species (ROS) increases oxidative stress and leads to cardiomyocyte apoptosis and necroptosis which is also defined as a programmed cell death. Purpose: In the present study, we investigate the effects of necrostatin-1 (Nec-1)—an inhibitor of receptor interaction proteins 1 (RIP1) and necroptosis—on doxorubicin-induced cardiotoxicity in rats. Research Design : Hearts were isolated and perfused by the Langendorff system in all four groups. Perfusion pressure (PP), left ventricular developed pressure (LVDP) and heart rate per minute (HR), LV (dP/dt) max, and LV (dP/dt) min which shows cardiac contractility and relaxation were recorded. Results: Results showed that PP significantly increased with Dox treatment and significantly decreased with Nec-1 treatment, while HR, LVDP, LV (dP/dt) max, and LV (dP/dt) min values significantly decreased with the Dox-treated group and significantly increased with Nec-1 treatment. Also with Nec-1 treatment, gene expression levels of anti-apoptotic Bcl-2 significantly increased and pro-apoptotic protein Bax, apoptotic marker caspase-3, and Nox-2 significantly decreased compared to the Dox-treated group. In heart tissues, MDA levels were significantly increased with Dox and decreased with Nec-1 treatment. These results were supported by the histological analysis indicated that Nec-1 reduced doxorubicin-induced cellular injury. Conclusions: In conclusion, our data indicate that Nec-1 ameliorates doxorubicin-induced cardiotoxicity by reducing oxidative stress injury and attenuating apoptosis and necroptosis. [ABSTRACT FROM AUTHOR]

Objective: To explore the antitumor effects of ethanol extract from Ventilago leiocarpa Benth (EEVLB) on sarcoma 180 (S180) tumor-bearing mice and the potential mechanism. Methods: Sixty mice were randomly assigned to 6 groups according to a random number table: normal group, model group, 5-fluorouracil (5-FU) group (0.02 g·kg−1), and high-, medium-, low-dose EEVLB groups (100, 84, and 56 g of raw material·kg−1 body weight, respectively), with 10 mice each group. All treatments were given once daily for 10 consecutive days. Effects of EEVLB on inhibiting tumor growth and immune function in mice were evaluated among all groups after the treatments by detecting tumor inhibition rate, organ index, serum levels of interleukin (IL)-2, -6, -10, CD3+CD4+ T lymphocytes, CD4+/CD8+ ratio, caspase-3 and Bcl-2. Results: EEVLB with different concentrations achieved inhibition of tumor growth in vivo, wherein the high-dose group showed the most significant reduction in tumor weight and increased apoptosis of tumor cells (P<0.05). In addition, both net weight gain and spleen index of mice showed uptrend in EEVLB treatment groups (P<0.05). Besides, serum levels of IL-2 and IL-6, percentages of CD3+CD4+ T lymphocytes and ratio of CD4+/CD8+ in peripheral blood were elevated in high- and medium-dose EEVLB groups compared with the model group (P<0.05). Also, upregulation of caspase-3 and downregulation of Bcl-2 were observed at protein levels in the high-dose EEVLB group (P<0.01). Conclusions: EEVLB exhibits promising antitumor activity in vivo. This effect might be due to activation of apoptotic signaling pathway, increase of cytokine levels and enhancement of immune function in tumor-bearing mice. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To investigate the pharmacodynamics of Rujikang naoemulsion preparation in the treatment of hyperplasia of mammary glands in mice. METHODS: The ICR mice were randomly divided into normal group, model group and treatment group [bromocriptine group（ coated with bromocriptine), high-dose group（ coated with high-dose Rujikang naoemulsion preparation） and low-dose group（ coated with low-dose Rujikang naoemulsion preparation） ]. The mouse model of mammary gland hyperplasia was established, after 30-day drug intervention on the treatment group, the changes in body mass, areola and nipple diameters were observed, the serum estradiol（ E2） and progesterone（ P） levels were measured, the changes in pathogeny structure of breast tissue were observed by hematoxylin-eosin staining method, the apoptotic cells were observed by TUNEL staining method, the Caspase-3 and Bcl-2 protein expression were detected by Western blot method. RESULTS: Compared with the model group, mice in the treatment group had lower body mass（ P < 0. 05）,shorter areola and nipple diameter（ P < 0. 01,P < 0. 001）,lower E2 level（ P < 0. 05）,higher P level（ P < 0. 05）,fewer in number of apoptotic cells（ P < 0. 001）,up-regulated Caspase-3 protein expression（ P < 0. 001）,and down-regulated Bcl-2 protein expression（ P < 0. 01）,differences were statistically significant. Compared with the model group, structures of mammary tissue and epithelial cell of the treatment group were basically restored to normal. CONCLUSIONS: The Rujikang naoemulsion preparation can significantly inhibit hyperplasia of mammary glands in mice caused by hormonal disorder. [ABSTRACT FROM AUTHOR]

Bhilawanol (Bh) and anacardic acid (AA) are two lipid-soluble compounds mostly found in the nut of Semecarpus anacardium (SA). This herb has many medicinal properties including enhancing learning and memory, yet its active compounds have not been studied for neuroprotective effects. We investigated the neuroprotective effects of Bh and AA against glutamate induced cell death in the adrenal pheochromocytoma cell line of rats (PC12 cells). Cell viability, toxicity and calcium influx were determined by MTT assay, LDH release assay and Fluo-3 imaging while apoptosis was assayed by caspase-3 and Bcl-2 gene expression. Our results showed that Bh and AA treatments significantly increased cell viability, reduced cell toxicity and calcium influx in PC12 cells in addition to suppressing the reactive oxygen species. Furthermore, AA treatment decreased caspase-3 expression level whereas both Bh and AA enhanced the expression of anti-apoptotic gene Bcl-2 in PC12 cells. Both compounds potently inhibited acetylcholinesterase enzyme (AChE) in a dose and time dependent manner. These findings suggest that the traditional use of SA may be explained on the basis of both Bh and AA showing neuroprotective potential due to their effects on enhancing cell viability, reducing cell toxicity most probably by reducing excessive calcium influx and suppression of ROS as well as by decreasing the expression of proapoptotic caspase 3 gene and increasing the expression of antiapoptotic gene Bcl2. Traditional use in enhancing learning and memory was justified in part by inhibition of AChE. [ABSTRACT FROM AUTHOR]

Bronchial asthma is a disease characterized by T-lymphocyte resistance to apoptosis. Recent studies have shown that there is an alternative form of cell death – autophagy. Since apoptosis in T-cells of patients with asthma is reduced, we suggested that autophagy could be initiated. The aim of the study is to perform a comparative analysis of the expression of apoptosis and autophagy key proteins in T-lymphocytes of patients with mild and severe bronchial asthma. To achieve this aim, the Western blot method was applied. The results were processed with the help of R statistics. Boxsplots were used to present the data. We showed that in the group with mild asthma the amount of caspase-3 increases during cultivation (spontaneous apoptosis is activated). In the group with the severe form, its content did not change. Autophagy activation was assessed by the presence of LC3-II protein. The results showed that form II of LC3-protein is present only in the group with the severe form. Therefore, the analysis of the content of key proteins of autophagy – Rubicon and UVRAG – in the cells of this category of patients was carried out and a significantly increased expression of these proteins was detected. In addition, we found a significant increase of UVRAG compared with Rubicon. Thus, inhibition of apoptosis is characteristic of T-lymphocytes of patients with bronchial asthma. However, in the mild group, a decrease of nutrients in the culture medium stimulated the launch of apoptosis, which was manifested in an increase in the content of caspase-3 and a decrease in Bcl-2. In the severe group, cell cultivation did not cause an increase in apoptotic activity, autophagy was activated instead, and we assume that for this group of patients it is an alternative way of cell death.

Breast cancer today is the most frequent cancer among women, and the second most common cause of cancer deaths among women. The aim of this study was to synthesize a new benzoxazole derivative, scan it for anti-cancer potential by MTT test using different breast cancer cell lines, and examine its effects on NF-κB and apopitosis-related proteins (APAF-1, cytochrome C, caspase-3, bcl-2) by the western blot method. newly-synthesized benzoxazole compound was applied to breast cancer cell lines (MDA-MB, MCF-7) and its cytotoxicity was measured quantitatively by MTT test. Later, the level of its effects on NF-κB and apopitosis-related proteins (APAF-1, cytochrome C, caspase-3, bcl-2) were examined by the western blot method. In our study, the structure of the synthesized new 5-[4-chlorobutanamido]-2-(p-methylphenyl)benzoxazole was proved by elemental analysis, 1H NMR and mass spectroscopy analysis methods. When the toxic effects of the application of the compound on the cell lines was examined by MTT, it had a greater toxic effect on MCF-7 when compared with MDA-MB, and IC50 levels were lower. When the protein was examined in immunohistochemistry with regard to VEGF, eNOS and TUNEL, it was observed that it caused a reduction in VEGF and an increase in eNOS and TUNEL. In the assay of the proteins by western blot, when benzoxazole compound was added to the MDA and MCF-7 cell line, there was no difference from the control group in Apaf-1 and BCL-2 levels, but a reduction was observed in caspase and Nfkβ levels compared with the control group. When the compound was added to the MDA-MB cell line, an increase was shown in the Cytochrome C level compared to the control group, but no difference was seen in the MCF-7 cell line. It is felt that this synthesized new benzoxazole compound increases apopitosis by reducing the activation of Nfkβ, and in this way has shown an effect of inhibiting tumor growth in cancer treatment. In addition, it is felt that this can provide hope in cancer treatment by the improved phase studies. [Med-Science 2019; 8(1.000): 186-91]

Background: Electroacupuncture (EA) pre-treatment can play a significant neuroprotective role in rats with cerebral ischaemia and reperfusion (CIR) injury, but the specific mechanism needs to be further elucidated. Objective: To investigate the effects of EA pre-treatment on apoptosis-related proteins Bax, Bcl-2, Cytochrome c (cyt c), cleaved caspase-9 and -3 in rats with CIR injury. Methods: CIR injury was induced using middle cerebral artery occlusion (MCAO) ischaemia-reperfusion. Thirty-six male Specific Pathogen Free (SPF) Sprague-Dawley rats were randomly divided into three groups: Sham (n=12), MCAO (n=12), and the EA+MCAO group (n=12). EA+MCAO group rats received electroacupuncture at DU20 for 2 consecutive weeks before MCAO model preparation. After 24 h of reperfusion, neurological deficits were evaluated by modified neurological severity scores. Brain infarct volumes were examined by 2,3,5-triphenyltetrazolium chloride staining. Expression of Bax, Bcl-2, cyt c, cleaved caspase-9 and -3 in the ischaemic penumbra were detected by Western blotting, and apoptosis evaluated by TUNEL staining. Results: After perfusion for 24 h, compared with the MCAO group, the neurological deficit scores and brain infarct volumes in the EA+MCAO group were significantly decreased (P <0.05), as was the level of Bax (P <0.05) or the Bax/Bcl-2 ratio (P <0.05), levels of cyt c, cleaved caspase-9 and -3 also decreased (P <0.05), like the number of TUNEL-positive cells (P <0.05), in contrast, the level of Bcl-2. Conclusion: EA pre-treatment exerts an anti-apoptotic effect through Bax-to-Bcl-2 ratio downregulation, blockage of mitochondrial cyt c release to the cytosol, and reduction of caspase-9 and -3 expression in rats with CIR injury. [ABSTRACT FROM AUTHOR]

Up-regulation of NF-κB and depletion of glutathione are acceptable mechanisms of Ifosfamide (IFO) renal toxicity. This investigation elucidates the role of free radicals, inflammatory and apoptotic markers in ifosfamide-induced rentoxicity and the protection of Annona species planted in Egypt (squamosa, cherimolia), and the hybrid between them (atemoya). Annona seeds extracts were prepared and phytochemical analyses were assessed. Rats were injected with saline or with IFO alone or in combination with Annona squamosa or Annona cherimolia or Annona atemoya orally. Biochemical, gene expression, histological and immune-histological examinations were performed. The results demonstrated that IFO elevated creatinine, sodium, magnesium and urea, along with depleted serum potassium and albumin levels. IFO caused a significant reduction in renal GSH, significant increases in renal MDA, and NOx, and up-regulated iNOS. In addition, IFO treatment showed increase mRNA and protein expression of NF-κB, while down-regulated mRNA and protein expression of Bcl-2, and strong immunohistological expression of caspase-3 as well as BAX in kidney tissues. Different Annona seed extracts significantly enhanced the sharpness of renal injury and improved oxidant responses induced by IFO. Annona extracts down-regulated iNOS, NF-κB expressions, as well as, down-regulated caspase-3 and BAX immunohistological expressions, and up-regulated Bcl-2 expression. A. squamosa showed a more pronounced protective effect. Histopathological examination was in accordance with biochemical results. Recent results suggest that Annona species ameliorated the renal toxicity of ifosfamide by down-regulation of NOx, iNOS, NF-κB, BAX, caspase-3 and MDA in addition to up-regulation of GSH and Bcl-2. [ABSTRACT FROM AUTHOR]

BACKGROUND: Reconstruction of the urinary tract with the ileum instead of the ureter has achieved a good clinical effect. There are yet no reports on what the adaptation relates to after the ileum acts as a substitute of the ureter. This study attempted to observe the effect of urine on the expression of apoptotic factors in intestinal epithelial cells, to further explore the possible underlying mechanism, and to provide experimental evidence for the pathological characteristics of the ileum instead of the ureter. OBJECTIVE: To observe the effect of urine environment on the expression of Caspase-3, Bax and Bcl-2 in human intestinal epithelial cells and to further investigate whether the urine environment promotes the apoptosis of intestinal epithelial cells and its underlying mechanism. METHODS: Human normal intestinal epithelial cells were treated with artificial urine that was diluted in culture medium at the rate of 0, 1:1, 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, and 1:128 for 24, 48, and 72 hours. Cell counting kit-8 was used to observe the effect of urine on the proliferation of intestinal epithelial cells. RTPCR and western blot assay were used to detect the mRNA and protein expressions of Caspase-3, Bax and Bcl-2 in intestinal epithelial cells. RESULTS AND CONCLUSION: Caspase-3 and Bax were highly expressed in intestinal epithelial cells of the group with the highest urine concentration, and the expression level gradually decreased as the urine concentration gradient decreased. The expression of Bcl-2 in intestinal epithelial cells was low in the group with the highest urine concentration, and the expression level gradually increased as the urine concentration gradient decreased. The higher the urine concentration, the higher the expression of pro-apoptotic factors Caspase-3 and Bax in intestinal epithelial cells, and the lower the expression of anti-apoptotic factor Bcl-2,indicating that the urine can promote the apoptosis of intestinal epithelial cells. [ABSTRACT FROM AUTHOR]

To determine the effect of green tea with the active ingredient epigallocathechin-3-gallate (EGCG) on the inhibition of apoptosis in the middle cerebral artery occlusion (MCAO) model. Four month old male Rattus norvegicus rats with a body weight of 200–275 g was used for the MCAO model and divided into five groups, and the treatment was carried out for 7 days. Before being sacrificed, the subject had 1 cc of blood drawn for high mobility group box 1 (HMGB-1) examination using enzyme-linked immunosorbent assay (ELISA), and after being sacrificed, the brain tissue specimen was taken to examine caspase-3 and B-cell lymphoma 3 (BCL-3) using immunohistochemistry methods. There was no significant difference in HMGB-1 results for the treatment group compared to the control group (P1: 384.20 ± 231.72 [p = 0.553]; P2: 379.11 ± 268.4 [p = 0.526]; P3: 284, 87 ± 276.19 [p = 0.140]; P4: 435.32 ± 279.95 [p = 0.912]). There is a significant increase in BCL-2 expression between the treatment group compared to the control group (P1: 2.58 ± 0.51 [p = 0.04]; P2: 3.36 ± 0.50 [p<0.001]; P3: 4.00 ± 0.42 [p<0.001]; P4: 3.60 ± 0.52 [p<0.001]). There was a significant difference in caspase-3 expression compared to the control group in the P3 group (P1: 4.33 ± 0.49 [p = 0.652]; P2: 4.09 ± 0.30 [p = 0.136]; P3: 3.58 ± 0.51 [p = 0.01]; P4: 3.89 ± 0.42 [p = 0.063]). There is no correlation between HMGB-1 and caspase-3 (r = −0.063; p = 0.613) or BCL-2 (r = −0.106; p = 0.396). There is significant negative correlation between caspase-3 and BCL-2 (r = −0.459; p = 0.000). Green tea with the active ingredient EGCG can inhibit neuronal cell death through the apoptotic pathway and not through the activation of HMGB-1. [ABSTRACT FROM AUTHOR]

Parkinson's disease (PD) is described as the loss of dopaminergic neurons located in the substantia nigra (SN) region of the brain and a progressive motor failure. Increased frequency of PD in women, especially after menopause, suggests the effect of estrogen. This view has been supported with empirical studies. Therefore, the effect of estrogen in an experimental model of Parkinson's disease induced by rotenone was investigated. A total of 32 female Wistar Albino rats were randomly assigned to four groups (control group, ovariectomy group, Parkinson's group, Parkinson's + estrogen group). The Parkinson's group received rotenone subcutanously at the dose of 2.5 mg/kg bw, on the 1st, 2nd, 3rd 4th, 6th, 9th, 12th, 15th, 18th, and 21st days animals in the Parkinson's + estrogen group received retonon as in the Parkinson's group and was additionally subcutaneously given estrogen (implant containing 0.5 mg 17 β-estradiol lasting for 21 days). The rats were subjected to rotarod, pole, and swimming tests at the end of the experiment for comparison of their motor activities, and then, histopathological and biochemical analyses were performed on the tissues that were extracted. The rotarod results revealed that Parkinson's group had the shortest time (32.33 ± 3.98 sn) than the groups of control (92.50 ± 12.60 s) ovariectomy (71.42 ± 10.58 s), and Parkinson's + estrogen (71.37 ± 9.26 s). The results of pole disclosed that return and landing time prolonged for Parkinson's group when compared with other groups (return time for control 2.98 ± 0.38 s, ovariectomy 3.02 ± 0.75 s, Parkinson 5.91 ± 0.33 s, Parkinson's + estrogen 3.48 ± 0.42 s and landing time for control 5.30 ± 0.59 s, ovariectomy 5.45 ± 0.73 s, Parkinson 9.80 ± 0.90 s, Parkinson's + estrogen 5.37 ± 1.02 s). Parkinson's group had longest (90.71 ± 12.56 s) swimming time to reach the target when compared with control (33.16 ± 8.68 s), ovariectomy (47.37 ± 12.19 s), and Parkinson's + estrogen (49.82 ± 5.78 s). Histopathological examination indicated a significant difference in tyrosine hydroxylase-stained cells (dopaminergic neurons and dopamine) between the Parkinson's + estrogen group and the Parkinson's group. The biochemical analyses of Caspas-3 activation in SN and striatum (STR) was significantly different between the Parkinson's + estrogen group and the Parkinson's group, but this difference was not observed in STR while evaluating Bcl-2. The results of this study suggested that estrogen may have a recuperative effect on PD. [ABSTRACT FROM AUTHOR]

AIM: To study the effect and mechanism of salvia ligustrazine on non-arteriti anterior ischemic optic neuropathy.METHODS: A total of 60 cases of non-arteriti anterior ischemic optic neuropathy were randomly divided into two groups. The control group(n=30 cases)was treated with oral prednisone acetate tablets and injection of compound anisodine. The experimental group(n=30 cases)was the same to the control group and added intravenous salvia ligustrazine, the period of treatment was 14d. The levels of IL-1β and TNF-α were detected by ELISA, and the protein expression levels of Bcl-2 and Caspase-3 were detected by Western blot. RESULTS: The best corrected visual acuity was improved in both groups. The plasma levels of IL-1β and TNF-α were significantly lowed in the experimental group than the control group, and the difference between the two groups was statistically significant(PPPCONCLUSION: Salvia ligustrazine promotes the recovery of visual function by reducing the level of inflammation and inhibiting cell apoptosis in body.

This study evaluated the potential of Punica granatum peel ethanol extract (PPEE) in attenuating the liver and kidney tissue injury induced by vancomycin (VM) treatment in rats. Fifty rats were distributed equally into five groups: control group, PPEE-administered group (100 mg/kg BW/day for 2 weeks; orally), VM-treated group (443.6 mg/kg BW, every alternate day for 2 weeks; intraperitoneally), pre-treated group, and concomitant-treated group. The biochemical response and the histopathology of the hepatic and renal tissue of the treated animals were assessed. The results showed that VM treatment induced substantial hepatotoxicity and nephrotoxicity, evidenced by a significant elevation in tissue injury and lipid oxidative (malondialdehyde) and inflammatory response (C-reactive protein) biomarkers, with lowered antioxidants and protein levels. Additionally, VM treatment induced various morphological, cytotoxic, vascular, and inflammatory perturbations as well as upregulation in the immune-expression of Caspase-3 and downregulation of BCL-2. Moreover, PPEE co-treatment was found to reduce the VM-induced toxicity by protecting the tissue against reactive oxygen species (ROS)–mediated oxidative damage, and inflammation as well as hinder the apoptotic cell death by modulating the expression of apoptosis-related proteins. Thus, we conclude that the PPEE administration showed more restoring efficacy when administered prior to VM medication. [ABSTRACT FROM AUTHOR]

Objective The models of delayed myocardial ischemia-reperfusion（I/R）injury in rats were constructed and verified by observing the apoptosis of myocardial cells and changes in cardiac function. Methods Forty-two SD rats were selected and suffered to the myocardial reperfusion after the anterior descending artery ligation for 1，4，6，12，and 24 h. The optimal ischemia time was selected with successful reperfusion appearance of anterior wall color and ST segment elevation recovery within 5 minutes. SD rats were divided into three groups including the sham operation group，timely reperfusion group，and delayed reperfusion group. Timely and delayed reperfusion cut-off time depended on the successful myocardial reperfusion appearance. Rats in the sham operation group were not ligated. The infarct area（IA)，risk area （AAR）and total ischemic area（IA+AAR）after reperfusion were measured and calculated by Evens Blue/TTC stainingat 24 h after reperfusion. Modified TUNEL assay was applied to detect the apoptosis of cardiomyocytes in the ischemic risk area of the three groups. The expression of apoptotic proteins Bax，Bcl-2 and cleaved Caspase-3 were detected by Western blotting at 24 h after reperfusion. The morphology and function indexes of the left ventricular end-systolic diameter （LVD)，left ventricular end-diastolic diameter（LVDd)，and left ventricular short-axis shortening rate（FS)，left ventricular end-systolic volume（LVDsV)，left ventricular end-diastolic volume（LVDdV)，left ventricular ejection fraction （LVEF)，ejection fraction per stroke（SS）and cardiac output（CO）were measured by the animal ultrasound machine at 72 h after reperfusion. Results Reperfusion was not achieved after 24 or 12 h of permanent ischemia. Only 2 of 10 rats were successfully perfused after 6 h of ischemia. One-hour and four-hour continuous ischemia were regarded as timely and delayed ischemia-reperfusion time，respectively. Compared with the timely reperfusion group，the necrotic area and ischemic area of the delayed reperfusion group increased（both P<0. 05），and the difference in the risk area was not statistically significant（P>0. 05）. The apoptosis rate in the delayed reperfusion group was higher than those in the timely reperfusion group and sham operation group（all P<0. 01）. Compared with the sham operation group，the expression of Bax and cleaved Caspase-3 increased significantly in both timely and delayed reperfusion groups（all P<0. 05 or P<0. 01），while Bcl-2 protein did not changed significantly. Compared with the sham operation group，FS，SS，LVEF and CO decreased in both delayed and timely reperfusion groups（all P<0. 01），and LVDsV and LVEF in the delayed reperfusion group were significantly lower than those in the timely reperfusion group（all P<0. 01）. Conclusions SD rats can be effectively reperfused after 4 h of myocardial ischemia，which can cause significant cardiomyocyte apoptosis and decrease left ventricular contractile function. This model can better simulate clinical delayed myocardial ischemia-reperfusion injury. [ABSTRACT FROM AUTHOR]

Objective To construct silk fibroin nanoparticles loaded with paclitaxel and to observe their effects on tumor growth and apoptosis protein expression in breast cancer bearing mice. Methods The silk fibroin was extracted from silk fiber，and then we added it to liquid paraffin for emulsification，glutaraldehyde for solidification，and obtained blank microspheres after freeze-drying. After paclitaxel was added to the ethanol solution to completely dissolve，the drugloaded microspheres were prepared according to the method of preparing blank microspheres. The breast cancer EMT6 cell suspension was injected subcutaneously into the right forelimb axillary of the mouse to establish the breast cancer-bearing mouse model. The 80 successfully modeled mice were randomly divided into the control group，paclitaxel group，paclitaxel carrier group，and blank microsphere group，with 20 mice in each. Mice in the paclitaxel group received 5 mg/kg intraperitoneal injection of paclitaxel on day 8，11，and 14 after vaccination. Mice in the paclitaxel carrier group received intraperitoneal injection of paclitaxel at the same time with a calculated concentration of 5 mg/kg drug-loaded microsphere solution. In the blank microsphere group，equal volume of blank microsphere solution was injected，and the mice in the control group were injected with an equal volume of normal saline. The concentration of paclitaxel in the tumor tissues was detected by high performance liquid chromatography（HPLC）method. The mouse orbital venous blood was collected and the serum ALT，AST，BUN，and creatinine levels were detected. The average tumor weight were recorded and tumor inhibition rate was calculated. Apoptosis rate of tumor cells was detected by flow cytometry. The expression of apoptosis-related proteins cleaved Caspase-3，Bax，and Bcl-2 in the tumor tissues were detected by Western blotting. Results The tumor growth inhibitory rate of the paclitaxel carrier group was higher than that of the paclitaxel group（P<0. 05）. No significant differences were found in the serum liver or kidney functions between the paclitaxel group，paclitaxel carrier group，blank microsphere group，and the control group（all P>0. 05）. The concentration of the paclitaxel carrier group was higher than that of the paclitaxel group（P<0. 05）. The apoptosis rates of paclitaxel carrier group and paclitaxel group were higher than those of the control group and blank microsphere group（all P<0. 05）. The tumor apoptosis rate of the paclitaxel carrier group was higher than that of the paclitaxel group（P<0. 05）. Compared with the control group and the blank microsphere group，the expression of cleaved Caspase-3 and Bax protein in the tumor tissues of the paclitaxel carrier group and the paclitaxel group increased，and the expression of Bcl-2 decreased（all P<0. 05）；the cleaved Caspase-3 and Bax protein expression in the paclitaxel carrier group was higher，and Bcl-2 expression was lower than those of the paclitaxel group（all P <0. 05）. Conclusion Silk fibroin nanoparticles loaded with paclitaxel can effectively inhibit tumor growth in mice with breast cancer，promote the apoptosis，and have a good anti-tumor effect，with no significant liver and kidney toxicity. [ABSTRACT FROM AUTHOR]

The impact of resveratrol (RSV) on radiation (RAD)‐induced brain injury in rats' brains was investigated. A total of 40 male Wistar Albino rats were randomly divided into four groups (control, RAD, RAD + RSV, and RSV groups, with 10 rats in each group). The results revealed a significant decrease in catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase activities, as well as glutathione (GSH) content. Further, a significant elevation in malondialdehyde, nitric oxide, interleukin‐1‐beta (IL‐1β), IL‐6, and transforming growth factor‐β1 levels were observed. Furthermore, decreased B‐cell lymphoma 2 (Bcl‐2), increased Bcl‐2‐associated X, and tumor necrosis factor‐α genes expression, decreased nuclear factor erythroid‐related factor 2, heme oxygenase‐1, and increased nuclear factor‐κB protein levels were noticed. Also, an apoptosis marker, caspase‐3‐positive cells, was seen in the hippocampus. Those effects were observed in the RAD group of rats. The treatment of RSV displayed a significant amendment of the studied parameters in the brain tissues of the RAD group of animals. This effect is interrelated to the ability of RSV to scavenge the free radicals, enhance the activity of the antioxidant enzymes, increase GSH contents, and downregulate the inflammatory responses and apoptosis markers in the brain tissues of RAD animals. In conclusion, this study demonstrated that the potent antioxidant, anti‐inflammatory, and antiapoptotic activities of RSV can improve the antioxidant status and suppress the inflammatory responses and apoptosis in the brain tissues of RAD animals. [ABSTRACT FROM AUTHOR]

BACKGROUND: Pinellia ternata can be cultivated artificially and is easy to purify. Its extract can inhibit the cell proliferation of chronic myeloid leukemia, acute myelogenous leukemia M3, and T-cell acute lymphoblastic leukemia, but the underlying mechanism remains unclear. OBJECTIVE: To explore the mechanism of action of pinellia ternata extract on apoptosis of three kinds of leukemia cells. METHODS: Chronic myeloid leukemia cell lines (K562), acute myeloid leukemia–M3 cell line (HL-60), acute T lymphocyte leukemia cell line (C8166) were incubated in five kinds of concentration in pinellia ternata extracts in vitro for 24, 48, and 72 hours, respectively. Cell counting kit-8 assay was used to assess the influence of three kinds of leukemia cell proliferation to screen the optimal inhibitory effect of moderate (300 mg/L) and high (500 mg/L) concentration. Flow cytometry was used to determine the incidence of early apoptosis of three kinds of leukemia cells. The mRNA and protein expression levels of Bax, Bcl-2 and Caspase-3 were detected by RT-PCR and western blot assay. RESULTS AND CONCLUSION: Cell counting kit-8 assay results showed that five different concentrations of pinellia extracts could inhibit the proliferation of three kinds of leukemia cells. The inhibition rates of medium concentration (300 mg/L) and high concentration (500 mg/L) were significantly higher than those of low concentration (100 mg/L) and control group (P < 0.01). Flow cytometry (Annexin PE/7AAD and Annexin V/PI) double staining showed that moderate and high concentrations of pinellia extract could induce early apoptosis of three kinds of leukemia cells. RT-PCR results revealed moderate and high concentrations of pinellia extract could down-regulate the mRNA expression level of Bcl-2, and up-regulate the mRNA expression levels of Bax, Bcl-2, Caspase-3 (P < 0.05, P < 0.01). Western blot assay results showed that the Bcl-2 protein bands of K562 and C8166 cells were significantly weakened after 72 hours of incubation with moderate and high concentrations of pinellia tubulin extract, while no significant changes were observed in HL-60 cells. Bax and caspase-3 protein bands were significantly enhanced. In summary, pinellia ternata extract can effectively inhibit the proliferation of myeloid leukemia and lymphocytic leukemia cells and promote their apoptosis via regulating Bax/Bcl-2 and Caspase-3 protein expression. [ABSTRACT FROM AUTHOR]

Objective(s): Chlorogenic acid is an herbal compound with various effects such as antiviral, antioxidant, and anticancer effect with low toxicity, which inhibits cell proliferation. Clinical studies had shown that chlorogenic acid has a positive effect on the different types of cancers treatment. Hence, this study evaluates chlorogenic acid effects on 4T1 breast cancer cells. Materials and Methods: In this study, cell proliferation was measured using an 3-(4,5-methylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay (MTT) on 4T1 cells. Afterwards, other assays like P53, Caspase-3 proteins expression and Annexin V/PI were detected by flow cytometry. Also; Bax and Bcl-2 were carried out by immunocytochemistry. Results: 200 μM of chlorogenic acid concentration showed the highest level of cytotoxicity toward 4T1 cells. Percentage of cell viability data were significant in 100 μM (P < 0.05) and 150, 200 μM (P < 0.001) doses. The evaluation using Annexin V/PI showed cell apoptosis in 100 μM (P < 0.05), 150 μM (P < 0.01), and for 200 μM (P < 0.05 and P < 0.01). Immunocytochemistry results showed the upregulation of Bax and also the downregulation of Bcl-2 in 4T1 cells treated with chlorogenic acid (P < 0.001). The expression level of P53 and caspase-3 increased during treatment with chlorogenic acid in the 4T1 cells (P < 0.001). Conclusion: Our findings demonstrated that chlorogenic acid plays a notable role on apoptosis inducing in the 4T1 cells through regulation of apoptotic proteins. [ABSTRACT FROM AUTHOR]

Trimethyltin chloride (TMT) is a stabilizer by-product in the process of manufacturing plastic, which is a kind of very strong toxic substance, and has acute, cumulative and chronic toxicity. TMT may cause bradycardia in patients with occupational poisoning, the mechanism of which has not been reported. This study explored the mechanism of TMT resulting in bradycardia of C57BL/6 mice. TMT was administered to mice to measure heart rate, serum succinate dehydrogenase (SDH) level, and myocardial Na+/K+-ATPase activity and expression. The effects of TMT on myocardial apoptosis were observed by changing the expressions of caspase-3, Bax and Bcl-2 in myocardium. It was found that the heart rate and SDH activity in serum of mice gradually decreased with the increase of TMT dose compared with the control group. The activity and the expression of Na+/K+-ATPase in the heart tissue of mice exposed to TMT was measured and gradually decreased with the increase of dose and time. We measured the expression of Bcl-2, Bax, caspase-3 and cleaved caspase-3 in the heart tissues of TMT exposed mice and found that the expressions of Bax, caspase-3 and cleaved caspase-3 increased and the expressions of Bcl-2 decreased in the heart tissues of the TMT-exposed mice at different doses. With the extension of TMT exposure time, the expression of Bax and caspase-3 increased and the expression of Bcl-2 decreased in the heart tissues of TMT exposed mice. Our findings suggest the mechanisms of TMT resulting in bradycardia may be associated with the inhibited activity and decreased content of Na+/K+-ATPase, thus further leading to the changes of Bcl-2, Bax, caspase-3 and cleaved caspase-3 in the mice's ventricular tissues. [ABSTRACT FROM AUTHOR]

The immunolocalization of apoptotic factors in rat neocortex was studied on the model of permanent occlusion of the middle cerebral artery with administration of exogenous BDNF. We revealed heterogeneous distribution of pro- and anti-apoptotic factors in the stroke area and in the surrounding penumbra, where caspase-3+ and p53+ cells were found. Their number was maximum on day 3 of ischemia. The number of neurons containing anti-apoptotic factors was relatively decreased. Injection of BDNF changed the distribution of apoptotic factors. In the penumbra area, BDNF enhanced the expression of Mdm2 primarily in the pyramid cells of layers V/VI and Bcl-2 in interneurons of layers II and III. Localization of p53 and caspase-3 varied at different terms of the ischemic period and showed an inverse dependence. Considering the selective neuroprotective effect of BDNF, various mechanisms of the formation of ischemic tolerance in neurons are proposed. [ABSTRACT FROM AUTHOR]

Introduction: Eyelid tumours mostly originated from skin and its appendeges. External carcinogens like UV radiation causes cell damages in the eyelid skin and contributes to carcinogenesis. Apoptosis is a very important mechanism to prevent these damage and probable neoplatic change. Aim: To compare caspase-3, p53 and Bcl-2 levels between patients with basal cell carcinoma (BCC) of the eyelid and healthy individuals. Material and methods: Pathology archives from October 2012 to April 2015 were scanned for BCC biopsies of the eyelid and tissue removed during blepharoplasty and entropion procedures. A total of 36 specimens were found. The specimens were divided into two groups: BCC group and controls (consisting of eyelid tissue removed during routine blepharoplasty). The pathology specimens were then stained using p53, Bcl-2 and caspase-3 stains and the intensity of staining was graded on a 0-3 scale. Results: Samples from a total of 36 patients were included in the study. Eighteen (50.0%) patients were female. There were 13 patients in the BCC group and 23 patients in the control group. The mean age was 66.0 ±10.8 years in the BCC group, and 65.61 ±11.22 years in the control group. The caspase-3 staining was lower in the BCC group than in the control group. No significant differences were found between the BCC group and the control group in terms of p53 levels or Bcl-2 levels (both of them, p = 1.000). Conclusions: The caspase-3 level was lower in the BCC group. This result suggests that these enzymes can play a significant role in carcinogenesis of eyelid BCC. [ABSTRACT FROM AUTHOR]