Search

Background: Existing studies have already revealed the involvement of C–C chemokine receptor type 7 (CCR7) in diverse human cancers, including esophageal cell squamous carcinoma (ESCA). Our current study, aims to explore the relevant mechanisms implicated. Methods: ESCA cell lines were collected for CCR7 expression quantification using western blot. Following the transfection, the viability, migration and invasion of ESCA cells were evaluated via cell counting kit-8 and Transwell assays. The specific molecular mechanisms underlying the effects of CCR7 in ESCA cells were explored via calculating the expressions of proteins related to metastasis and Janus kinase 2/signal transduction and transcription activation 3 (JAK2/STAT3) signaling pathway via western blot. The correlation between CCR7 and metastasis-related proteins was explored via Pearson's correlation test. Results: CCR7 was high-expressed in ESCA cells and CCR7 knockdown repressed the viability, migration and invasion of ESCA cells, concurrent with the increased expression of E-cadherin (E-cad, which was also known as CDH1 and lowly expressed in ESCA cells) and the decreased expressions of vimentin (Vim, which was highly expressed in ESCA cells) and matrix metalloproteinase-9 (MMP-9, which was also highly expressed in ESCA cells). Meanwhile, CCR7 was positively correlated with Vim and MMP-9 yet negatively correlated with E-cad in ESCA cells, which indicated that CCR7 has a role in promoting tumor progression in ESCA cells. Besides, the phosphorylation of STAT3 and JAK2 in ESCA cells was elevated, which was diminished following CCR7 knockdown. Conclusion: This study proves the modulation of CCR7 on ESCA in vitro, which was achieved via JAK2/STAT3 signaling pathway. Our discovery will provide new therapeutic basis and insights for ESCA. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of dexmedetomidine (DEX) on the polarization of alveolar macrophages induced by lipopolysaccharide (LPS) and to explore the related mechanisms. Methods: Rat alveolar macrophages NR8383 were cultured in vitro. Experiment one was divided into control group, model group (1 μg/ml LPS), DEX low, medium and high dose groups (1, 5, 10 mg/kg DEX+10 mg/kg LPS). Experiment two was divided into DEX high dose group (10 mg/kg) and DEX high dose+Colivelin (JAK2/STAT3 signaling pathway activator) group (10 mg/kg DEX+0.5 μmol/L Colivelin). The morphological changes of rat alveolar macrophages NR8383 were observed by inverted microscope; RT-PCR method was used to detect the expression levels of iNOS and Arg1 mRNA in NR8383 cells, and flow cytometry was used to detect the expression levels of CD86 and CD163 proteins in NR8383 cells; Western blot was used to detect the expression levels of surface marker proteins TNF-α, iNOS, SOCS, Arg1, TGF-β and JAK2/ STAT3 signaling pathway related proteins in NR8383 cells. Results: Compared with control group, there were a lot of cell debris in the intercellular space of NR8383 in the model group, the proportions of iNOS mRNA, CD86 positive cells, and the expression levels of TNF-α, p-JAK2/JAK2, p-STAT3/STAT3 were significantly increased, the proportions of Arg1 mRNA, CD163 positive cells, and the expression levels of SOCS and TGF-β were significantly reduced (P<0.05); compared with the model group, the NR8383 intercellular cell debris in the DEX low, medium, and high dose groups were decreased, the proportions of iNOS mRNA, CD86 positive cells, and the expression levels of TNF-α, p-JAK2/JAK2, p-STAT3/STAT3 were significantly reduced, the proportions of Arg1 mRNA, CD163 positive cells, and the expression levels of SOCS and TGF- β were significantly increased (P<0.05). The reactivation of the JAK2/ STAT3 signal pathway by Colivelin could weaken the role of DEX in LPS induced NR8383 cell polarization. Conclusion: DEX can inhibit the M1 polarization of NR8383 cells induced by LPS, which may be achieved by inhibiting the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

AbstractObjective: This study aims to explore the effects of Triptolide (TP) on the differentiation of Th17 cells in ankylosing spondylitis (AS).Methods: Peripheral blood mononuclear cells (PBMCs) collected from 10 patients with active AS patients were exposed to TP, GSK-J4 or vehicle. T lymphocyte subsets were analyzed using flow cytometry. ELISA was used to assess the level of IL-17. Western blot analysis and quantitative RT–PCR were used to measure the mRNA and protein levels of RORγt, JMJD3, EZH2, JAK2 and STAT3 in the JAK2/STAT3 signaling pathway.Results: We observed a tendency toward a greater percentage of IL-17-positive CD4+ T cells in peripheral blood mononuclear cells (PBMCs) from patients with active AS than in those from healthy controls. Triptolide (TP) and GSK-J4 significantly reduced IL-17 expression. In cultured PBMCs from patients with active AS, 24 h of treatment with TP or GSK-J4 decreased the expression of RORγt (p < 0.05), JAK2 and STAT3 (JAK2: p < 0.05; STAT3: p < 0.05). Furthermore, both triptolide and GSK-J4 increased the level of histone 3 with Lys 27 trimethylation (H3K27me3) in patient-derived PBMCs. H3K27me3 enrichment was detected at the promoters of the RORc, STAT3 and IL-17 genes. Consistent with this finding, triptolide upregulated the EZH2 gene and downregulated the JMJD3 gene.Conclusion: Triptolide inhibits Th17 cell differentiation via H3K27me3 upregulation and orchestrates changes in histone-modifying enzymes, including JMJD3 and EZH2. These findings support the clinical efficacy of triptolide for AS and may provide clues for identifying molecular targets for the development of novel treatments. [ABSTRACT FROM AUTHOR]

To investigate the effects of resveratrol (Res) on human fetal scleral fibroblasts (HFSFs) and its potential mechanism. HFSFs were randomly divided into the Res-treated group and the control group. Following, HFSFs were treated with or without a concentration of 10 μM Res for 48 h. To detect the expression of related genes, reverse transcription quantitative PCR (RT-qPCR) and western blotting were used. The apoptosis rate of different groups was determined using flow cytometry. The mRNA expression of matrix metalloproteinase 2 (MMP-2), Collagen, Type I, Alpha 1 (COL1A1), Janus Kinase 2 (JAK2), and Signal Transducer and Activator of Transcription 3 (STAT3)" was downregulated in the Res-treatment group compared to the control group, according to RT-qPCR. Western blotting revealed that Res therapy reduced the expression of MMP-2, JAK2, P-JAK2, STAT3, P-STAT3, and Bcl-2 associated protein X (Bax) while increasing the expression of COL1A1 and B-cell lymphoma-2 (Bcl-2). Flow cytometry showed that the cell apoptosis rate was significantly lower in HFSFs treated with Res. In conclusion, these findings suggest that Res increases COL1A1 expression while inhibiting MMP-2 and cell apoptosis in HFSFs, possibly through modulation of the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

Abstract Background Existing studies have already revealed the involvement of C–C chemokine receptor type 7 (CCR7) in diverse human cancers, including esophageal cell squamous carcinoma (ESCA). Our current study, aims to explore the relevant mechanisms implicated. Methods ESCA cell lines were collected for CCR7 expression quantification using western blot. Following the transfection, the viability, migration and invasion of ESCA cells were evaluated via cell counting kit-8 and Transwell assays. The specific molecular mechanisms underlying the effects of CCR7 in ESCA cells were explored via calculating the expressions of proteins related to metastasis and Janus kinase 2/signal transduction and transcription activation 3 (JAK2/STAT3) signaling pathway via western blot. The correlation between CCR7 and metastasis-related proteins was explored via Pearson’s correlation test. Results CCR7 was high-expressed in ESCA cells and CCR7 knockdown repressed the viability, migration and invasion of ESCA cells, concurrent with the increased expression of E-cadherin (E-cad, which was also known as CDH1 and lowly expressed in ESCA cells) and the decreased expressions of vimentin (Vim, which was highly expressed in ESCA cells) and matrix metalloproteinase-9 (MMP-9, which was also highly expressed in ESCA cells). Meanwhile, CCR7 was positively correlated with Vim and MMP-9 yet negatively correlated with E-cad in ESCA cells, which indicated that CCR7 has a role in promoting tumor progression in ESCA cells. Besides, the phosphorylation of STAT3 and JAK2 in ESCA cells was elevated, which was diminished following CCR7 knockdown. Conclusion This study proves the modulation of CCR7 on ESCA in vitro, which was achieved via JAK2/STAT3 signaling pathway. Our discovery will provide new therapeutic basis and insights for ESCA.

BACKGROUND: Acupuncture is an effective method for lumbar pain in lumbar disc herniation, but its mechanism has not yet been clarified. Factors related to the JAK2/STAT3 signaling pathway regulate the body’s inflammatory response and are involved in the process of neuropathic pain. OBJECTIVE: To study the mechanism of acupuncture on lumbar disc herniation in a rat model based on the JAK2/STAT3 signaling pathway. METHODS: Forty Sprague-Dawley rats were randomly divided into four groups: sham operation group, model group, acupuncture group, and acupuncture+agonist group, with 10 rats in each group. Animal models of L5 lumbar disc herniation was constructed through autologous disc cell transplantation in the model group, acupuncture group, and acupuncture+agonist group. Rats in the acupuncture group and the acupuncture+agonist group received acupuncture treatment (Yanglingquan, Shenshu, Huantiao, and Dachangshu acupoints) at 3 days after modeling, and acupuncture treatment was given once a day, 20 minutes each, for 15 consecutive days. Rats in the acupuncture+agonist group were injected intrathecally with coumermycin A1, a JAK2 agonist, into the L4/L5 intervertebral space, once a day, 20 minutes each, prior to the acupuncture at 6, 12, and 18 days after modeling. Paw withdrawal mechanical threshold was detected before and 3, 6, 9, 12, 15, and 18 days after modeling. At 18 days after modeling, serum inflammatory factor levels were detected, hematoxylin-eosin staining was performed to observe the morphology of L5-L6 tissues, RT-PCR was performed to detect the expression of JAK2 and STAT3 mRNAs in L5-L6 tissues, and western blot was performed to detect the expression of JAK2, p-JAK2 and p-STAT3 proteins in L5-L6 tissues. RESULTS AND CONCLUSION: The paw withdrawal mechanical thresholds of rats in the model group at different time points after modeling were lower than those in the sham operation group (P < 0.05), the paw withdrawal mechanical thresholds of rats in the acupuncture group were higher than those in the model group at 9, 12, 15, and 18 days after modeling (P < 0.05), and the paw withdrawal mechanical thresholds of rats in the acupuncture+agonist group were lower than those in the acupuncture group at 9, 12, 15, and 18 days after modeling (P < 0.05). The levels of interleukin 6, tumor necrosis factor α, neurotransmitter substance P, and brain neuropeptide Y were elevated in the model group compared with the sham operation group (P < 0.05); the levels of all four inflammatory factors were reduced in the acupuncture group compared with the model group (P < 0.05); and the levels of all four inflammatory factors were elevated in the acupuncture+agonist group compared with the acupuncture group (P < 0.05). Hematoxylin-eosin staining showed that lumbar degeneration was obvious in the model group but reduced in the acupuncture group and the acupuncture+agonist group. Moreover, the reduction was more obvious in the acupuncture group compared with the acupuncture+agonist group. The JAK2 and STAT3 mRNA expression as well as the p-JAK2 and p-STAT3 protein expression were elevated in the model group compared with the sham operation group (P < 0.05), were decreased in the acupuncture group compared with the model group (P < 0.05), and were increased in the acupuncture+agonist group compared with the acupuncture group (P < 0.05). To conclude, acupuncture can alleviate inflammation to exert analgesic effects in the rat model of lumbar disc herniation, and its mechanism of action may be related to the inhibition of the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

Abstract To explore the effects and mechanisms of the Xianhecao-Huanglian drug pair on autophagy-mediated intervention in acute inflammatory bowel disease (IBD) via the JAK2/STAT3 pathway. The study examined the underlying mechanisms of action of Xianhecao (APL) and Huanglian (CR) using a mouse model of dextran sodium sulfate (DSS)-induced acute inflammatory bowel disease (IBD) and in an in vitro model of IBD induced by lipopolysaccharide (LPS). The assessment of the therapeutic efficacy of the Xianhecao-Huanglian drug combination in a mouse model of IBD caused by DSS included the following parameters: Assessment of weight loss or gain. Measurement of the disease activity index (DAI). Assessment of histological damage. Determination of organ index. Measurement of colon length. Ascertain the levels of inflammatory cytokines in the intestinal tissues and serum of mice. Immunohistochemistry (IHC) for the measurement of tight junction protein concentrations in the colon mucosa, including ZO-1, claudin-1, and occludin. Measurement of mucin levels, specifically Mucin 2 (Muc2). Hematoxylin and eosin (HE) staining for the observation of histopathological alterations in colonic tissues. Examining the effect on goblet cells using periodic acid-Schiff (PAS) labeling. Application of Western blot and immunofluorescence techniques for the detection of autophagy-related markers in colonic tissues and proteins associated with the JAK2/STAT3 pathway. A cell inflammation model of IBD was induced through LPS stimulation, and a serum containing the Xianhecao-Huanglian drug pair (referred to as ACHP-DS) was formulated. Cell viability, anti-proinflammatory cytokines, tight junction proteins, mucins, autophagy-related markers, and the JAK2/STAT3 signaling pathway were assessed. The Xianhecao-Huanglian drug pair significantly ameliorated the symptoms and survival quality of acute IBD mice, reducing the disease activity index score, raising MUC2 secretion and tight junction protein expression to improve the integrity of the intestinal barrier, and preserving goblet cell function; thus, protecting the intestines. It effectively restrained triggering the signaling pathway that involves JAK2 and STAT3, leading to the suppression of inflammation and amelioration of colonic inflammation damage. Additionally, it induced autophagy in mouse colonic tissues.The in vitro experiments demonstrated that the Xianhecao-Huanglian drug combination enhanced the viability of LOVO and NCM460 cells when exposed to LPS stimulation. Furthermore, it suppressed the production of inflammatory cytokines such as IL-6, IL-1β, as well as TNF-α, whilst increasing the production of IL-10, ZO-1, along with MUC2. These effects collectively led to the alleviation of inflammation and the restoration of mucosal integrity. The results were consistent with what was shown in the in vivo trial. Moreover, the medication demonstrated effectiveness in reducing JAK2 along with STAT3 phosphorylation levels in the LPS-induced inflammatory model of IBD cells. The intervention with either the Xianhecao-Huanglian drug combination-containing serum or the JAK2/STAT3 pathway inhibitor AG490 reversed the pro-inflammatory effects and increased autophagy levels in the LPS-stimulated cells. The Xianhecao-Huanglian drug combination modulates the JAK2/STAT3 pathway, leading to the induction of autophagy, which serves as an intervention for IBD.

ObjectiveTo investigate the effect of folic acid–modified liposome quercetin (FLQ) on the proliferation and apoptosis of triple negative breast cancer (TNBC) cells and explore its underlying mechanism. MethodsCCK-8 was used to detect the effect of FLQ on TNBC cell viability. Colony formation assay was conducted to detect the effect of FLQ on TNBC cell proliferation. Flow cytometry was performed to detect the effect of FLQ on TNBC cell apoptosis, the levels of intracellular ROS, and mitochondrial membrane potential. Western blot analysis was conducted to detect the expression levels of JAK2/STAT3 signaling pathway-related and apoptosis-related proteins. ResultsFLQ inhibited the proliferation and promoted the apoptosis of MDA-MB-231 cells (P=0.023, P

To explore the effects and mechanisms of the Xianhecao-Huanglian drug pair on autophagy-mediated intervention in acute inflammatory bowel disease (IBD) via the JAK2/STAT3 pathway. The study examined the underlying mechanisms of action of Xianhecao (APL) and Huanglian (CR) using a mouse model of dextran sodium sulfate (DSS)-induced acute inflammatory bowel disease (IBD) and in an in vitro model of IBD induced by lipopolysaccharide (LPS). The assessment of the therapeutic efficacy of the Xianhecao-Huanglian drug combination in a mouse model of IBD caused by DSS included the following parameters: Assessment of weight loss or gain. Measurement of the disease activity index (DAI). Assessment of histological damage. Determination of organ index. Measurement of colon length. Ascertain the levels of inflammatory cytokines in the intestinal tissues and serum of mice. Immunohistochemistry (IHC) for the measurement of tight junction protein concentrations in the colon mucosa, including ZO-1, claudin-1, and occludin. Measurement of mucin levels, specifically Mucin 2 (Muc2). Hematoxylin and eosin (HE) staining for the observation of histopathological alterations in colonic tissues. Examining the effect on goblet cells using periodic acid-Schiff (PAS) labeling. Application of Western blot and immunofluorescence techniques for the detection of autophagy-related markers in colonic tissues and proteins associated with the JAK2/STAT3 pathway. A cell inflammation model of IBD was induced through LPS stimulation, and a serum containing the Xianhecao-Huanglian drug pair (referred to as ACHP-DS) was formulated. Cell viability, anti-proinflammatory cytokines, tight junction proteins, mucins, autophagy-related markers, and the JAK2/STAT3 signaling pathway were assessed. The Xianhecao-Huanglian drug pair significantly ameliorated the symptoms and survival quality of acute IBD mice, reducing the disease activity index score, raising MUC2 secretion and tight junction protein expression to improve the integrity of the intestinal barrier, and preserving goblet cell function; thus, protecting the intestines. It effectively restrained triggering the signaling pathway that involves JAK2 and STAT3, leading to the suppression of inflammation and amelioration of colonic inflammation damage. Additionally, it induced autophagy in mouse colonic tissues.The in vitro experiments demonstrated that the Xianhecao-Huanglian drug combination enhanced the viability of LOVO and NCM460 cells when exposed to LPS stimulation. Furthermore, it suppressed the production of inflammatory cytokines such as IL-6, IL-1β, as well as TNF-α, whilst increasing the production of IL-10, ZO-1, along with MUC2. These effects collectively led to the alleviation of inflammation and the restoration of mucosal integrity. The results were consistent with what was shown in the in vivo trial. Moreover, the medication demonstrated effectiveness in reducing JAK2 along with STAT3 phosphorylation levels in the LPS-induced inflammatory model of IBD cells. The intervention with either the Xianhecao-Huanglian drug combination-containing serum or the JAK2/STAT3 pathway inhibitor AG490 reversed the pro-inflammatory effects and increased autophagy levels in the LPS-stimulated cells. The Xianhecao-Huanglian drug combination modulates the JAK2/STAT3 pathway, leading to the induction of autophagy, which serves as an intervention for IBD. [ABSTRACT FROM AUTHOR]

BACKGROUND: Shujin Jiannao Prescription is an empirical formula for the treatment of cerebral palsy in Dongzhimen Hospital, Beijing University of Chinese Medicine, with clear clinical efficacy, but the specific mechanism needs to be elucidated. OBJECTIVE: To explore the possible mechanism of Shujin Jiannao Prescription in treating cerebral palsy. METHODS: Sixty-four 7-day-old Sprague-Dawley rats were randomly divided into a normal group (n=12) and a model group (n=52). An animal model was established by the Rice-Vannucci method. After successful modeling, 52 model rats were randomly divided into control model group (n=12), minocycline group, and the low-, medium-, and high-dose groups of the Shujin Jiannao Prescription (n=10 per group). Rats in the minocycline group were given 40 mg/kg•d minocycline by gavage; rats in the low-, medium, and high-dose groups were given 4, 8, and 16 g/kg•d Shujin Jiannao Prescription granules by gavage, respectively; and rats in the normal group and control model group were given an equal dose of normal saline by gavage. Medication in each group was given once a day for 1 week. The rats in each group were evaluated behaviorally using suspension test, abnormal involuntary movement score, and Bederson score. The pathological changes of the cerebral cortex were observed by hematoxylin-eosin staining. The levels of tumor necrosis factor α, interleukin 1β, and interleukin 10 in the cerebral cortex were determined using ELISA. The positive expressions of Janus kinase 2 (JAK2), phosphorylated Janus kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3) in the cerebral cortex were detected using immunohistochemistry. The protein expression levels of JAK2, p-JAK2, and p-STAT3 were detected using western blot. RESULTS AND CONCLUSION: Compared with the normal group, the suspension test score and involuntary movement score were decreased in the control model group (P < 0.01 or P < 0.05). The pathological results showed structural disruption of nerve cells, formation of large numbers of vacuoles, cell swelling, and increased intercellular space in the control model group. In addition, the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were significantly increased (P < 0.01), the expression of interleukin 10 was decreased (P < 0.05), and the protein expressions of JAK2, p-JAK2, and p-STAT3 in the cerebral cortex were significantly increased (P < 0.01) in the control model group compared with the normal group. Compared with the model group, minocycline and Shujin Jiannao Prescription at each dose could improve the behavioral indexes of rats (P < 0.01 or P < 0.05) and ischemic-hypoxic pathological changes were attenuated, with only a small amount of necrotic nerve cells and a few vacuoles, and reduced intercellular space. Moreover, the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were decreased in each drug group compared with the control model group (P < 0.05), while the protein expressions of JAK2, p-JAK2, and p-STAT3 in the cerebral cortex were significantly decreased (P < 0.01). The most obvious improvement was observed in the high-dose Shujin Jiannao Prescription group. To conclude, Shujin Jiannao Prescription can inhibit inflammation in the cerebral cortex of rats with hypoxic-ischemic brain injury. The mechanism may be related to the regulation of the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

Background: Oleanolic acid (OA), a pentacyclic triterpenoid exhibiting specific anti-cancer properties and highly effective antioxidant activity, was isolated from traditional Chinese medicinal herbs. Conversely, the OA that impacts colon cancer (CC) cells and its underlying mechanisms remain poorly understood. Methods: The cytotoxic effect of OA alone or OA-5-Fluorouracil (5-FU) combination on normal and CC cells was analyzed by methyl thiazolyl diphenyl-tetrazolium bromide (MTT). Then, the impact of OA on CC cell lines (LoVo and HT-29) proliferation and stemness were measured using colon formation and tumorsphere formation assays. Octamerbinding transcription factor 4 (Oct4), Prominin-1 (CD133), Nanog, and transcription factor SOX-2 (SOX2) are cell stemness-related indicators whose expression was assessed using fluorescence qPCR assay, Western blotting, and immunohistochemistry. The effect of OA on the proliferative potency of CC cells was evaluated using an in vivo model. Results: The stem-like characteristics and clone production of colon cancer cells were markedly reduced by OA alone or in combination with OA-5-FU. Moreover, OA increases the susceptibility of CC cells to 5-FU by blocking the cell stemness-related markers (CD133, Nanog, SOX2, and Oct4) expression levels both in vitro and in vivo, as well as by inactivating the activator of transcription 3 (STAT3 signaling) and Janus kinase 2/signal transducer (JAK2). Conclusion: These findings imply that oleanolic acid, both in vitro and in vivo, suppresses the JAK2/STAT3 pathway, which in turn reverses chemoresistance and decreases colon cancer cell stemness. Therefore, by reducing the recommended amount of 5-FU, this strategy may improve chemotherapeutic effectiveness and minimize undesired side effects. [ABSTRACT FROM AUTHOR]

Objective To investigate the effect of folic acid–modified liposome quercetin (FLQ) on the proliferation and apoptosis of triple negative breast cancer (TNBC) cells and explore its underlying mechanism. Methods CCK-8 was used to detect the effect of FLQ on TNBC cell viability. Colony formation assay was conducted to detect the effect of FLQ on TNBC cell proliferation. Flow cytometry was performed to detect the effect of FLQ on TNBC cell apoptosis, the levels of intracellular ROS, and mitochondrial membrane potential.Western blot analysis was conducted to detect the expression levels of JAK2/STAT3 signaling pathway-related and apoptosis-related proteins. Results FLQ inhibited the proliferation and promoted the apoptosis of MDA-MB-231 cells (P=0.023, P<0.001).It promoted mitochondrial membrane potential collapse and increased the intracellular ROS levels of MDA-MB-231 cells (P=0.003, P=0.034); inhibited the phosphorylation levels of JAK2 and STAT3; upregulated the expression levels of the proapoptotic proteins Bax, Bak, cytochrome C, and Cleaved-Caspase-3 (P<0.001, P<0.001); and downregulated the expression levels of the antiapoptotic proteins Bcl2 and Bcl-xL (P=0.037, 0.028). Conclusion FLQ inhibits the proliferation and induces the apoptosis of MDA-MB-231 cells. These effects may be related to the activation of the mitochondrial apoptosis pathway through the inhibition of the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

Adenomyosis (ADS) is a common benign gynecological disease. Abnormal proliferation at the endometrial-myometrial interface (EMI) plays a crucial role in the occurrence and progression of ADS. miR-141-3p is associated with cell proliferation and apoptosis. However, the specific mechanism of miR-141-3p in the etiology of ADS is still unknown. In this study, we explored the effects of miR-141-3p on the proliferation and apoptosis of ADS EMI smooth muscle cells (SMCs). We collected EMI tissues for the primary culture of SMCs from 25 patients diagnosed with ADS and 20 without ADS. Real-time quantitative polymerase chain reaction and western blot were used to measure the mRNA and protein expression levels of miR-141-3p, JAK2, STAT3, phospho-JAK2, and phospho-STAT3 in ADS EMI SMCs. The cell counting kit 8 assay and flow cytometry analysis were used to evaluate the proliferation and apoptosis of EMI SMCs. The miR-141-3p mimic/inhibitor was used to increase or decrease the expression level of miR-141-3p. We added WP1066 to block the phosphorylation of JAK2/STAT3 pathway components. The miR-141-3p levels were decreased, while JAK2 and STAT3 levels were increased in ADS EMI SMCs. miR-141-3p overexpression significantly inhibited the proliferation and enhanced the apoptosis of EMI SMCs, whereas a decrease in miR-141-3p expression level was connected to the opposite results. Meanwhile, inactivated JAK2/STAT3 pathway decreased proliferation and enhanced apoptosis of EMI SMCs after WP1066 treatment. Furthermore, rescue experiments confirmed that the JAK2/STAT3 pathway was the downstream pathway of miR-141-3p and reduced the effect of miR-141-3p on the proliferation and apoptosis of EMI SMCs. These results demonstrate that miR-141-3p regulates the proliferation and apoptosis of ADS EMI SMCs by modulating the JAK2/STAT3 pathway. [ABSTRACT FROM AUTHOR]

Diabetic cardiomyopathy (DCM) represents a common pathological state brought about by diabetes mellitus (DM). Patchouli alcohol (PatA) is known for its diverse advantageous effects, notably its anti-inflammatory properties and protective role against metabolic disorders. Despite this, the influence of PatA on DCM remains relatively unexplored. To explore the effect of PatA on diabetes-induced cardiac injury and dysfunction in mice, streptozotocin (STZ) was used to mimic type 1 diabetes in mice. Serological markers and echocardiography show that PatA treatment protects the heart against cardiomyopathy by controlling myocardial fibrosis but not by reducing hyperglycemia in diabetic mice. Discovery Studio 2017 software was used to perform reverse target screening of PatA, and we found that JAK2 may be a potential target of PatA. RNA-seq analysis of heart tissues revealed that PatA activity in the myocardium was primarily associated with the inflammatory fibrosis through the Janus tyrosine kinase 2 (JAK2)/signal transducer and activator of the transcription 3 (STAT3) pathway. In vitro, we also found that PatA alleviates high glucose (HG) + palmitic acid (PA)-induced fibrotic and inflammatory responses via inhibiting the JAK2/STAT3 signaling pathway in H9C2 cells. Our findings illustrate that PatA mitigates the effects of HG + PA- or STZ-induced cardiomyopathy by acting on the JAK2/STAT3 signaling pathway. These insights indicate that PatA could potentially serve as a therapeutic agent for DCM treatment. [ABSTRACT FROM AUTHOR]

Osteoporosis (OP) is a common chronic progressive bone disease that increases fracture risk in postmenopausal women. Research suggests that puerarin (Pue) may be an effective treatment for OP. This study examined the effects and underlying mechanisms of Pue in treating postmenopausal osteoporosis (PMOP) in rats. Sprague–Dawley (SD) rats underwent bilateral ovariectomy to simulate PMOP and were then treated with subcutaneous injections of Pue. Bone mineral density (BMD) was measured using a bone densitometer. Micro-CT scans assessed femur bone structure and various parameters were calculated: bone volume fraction (BV/TV), bone surface density (BS/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular separation (Tb.Sp), and bone surface area-to-bone volume ratio (BS/BV). Hematoxylin-eosin (HE) staining was employed to observe femoral tissue pathology. Serum levels of bone formation metabolism-related markers—osteocalcin (OC), bone alkaline phosphatase (BALP), and procollagen type I N-terminal propeptide (PINP)—were measured via enzyme-linked immunosorbent assay (ELISA). The protein expression levels of the Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway in bone tissue were evaluated using Western blotting assay. The results showed improved bone density and reduced bone loss in rats treated with Pue. There were also significant increases in serum levels of OC and BALP, indicating enhanced osteogenesis. Furthermore, there was a decrease in activation of the JAK2/STAT3 pathway in femoral tissue, suggesting a pathway inhibition. These findings indicate that Pue may combat osteoporosis by promoting osteogenesis and inhibiting activation of the JAK2/STAT3 pathway activation.

Objective To investigate the influence of limonin on the malignant biological behavior of non-small cell lung cancer (NSCLC) cells by regulating the protein tyrosine kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Methods CCK-8 method was applied to detect the survival rate of A549 cells treated with different concentrations of limonin (0, 5, 10, 25, 50, 75, 100 μmol/L). A549 cells were separated into normal culture (NC) group, low-dose limonin group (treatment with 10 μmol/L limonin for 24 h), medium-dose limonin group (treatment with 25 μmol/L limonin for 24 h), high-dose limonin group (treatment with 50 μmol/L limonin for 24 h), coumermycin A1 group (treatment with 10 μmol/L JAK2 activator coumermycin A1+50 μmol/L limonin for 24 h), and AG490 group (treatment with 10 μmol/L JAK2 inhibitor AG490+50 μmol/L limonin for 24 h). Clone formation assay was applied to detect the clones of each group of cells. Transwell assay was applied to detect cell migration and invasion, and flow cytometry was applied to detect apoptosis. Western blot analysis was applied to detect the protein expression levels of JAK2, p-JAK2, STAT3, p-STAT3, E-cadherin, N-cadherin, and vimentin in each group. Results The viability of A549 cells decreased significantly in a limonin concentration-dependent manner (P < 0.05), with IC50 of 45.16±1.66 μmol/L. Concentrations of 10, 25, and 50 μmol/L were selected for subsequent experiments. The numbers of clones, migration, and invasion of A549 cells and the protein expression levels of IL-6, p-JAK2, p-STAT3, N-cadherin, and vimentin in the low-, medium-, and high-dose limonin groups significantly decreased, compared with those in the NC group, and the apoptosis rate and E-cadherin protein expression significantly increased (P < 0.05). The JAK2 activator coumermycin A1 attenuated the ability of limonin to inhibit the proliferation, migration, invasion, and other malignant biological behavior of A549 cells and attenuated the apoptosis ability. The JAK2 inhibitor AG490 enhanced the ability of limonin to inhibit the proliferation, migration, invasion, and other malignant biological behavior of A549 cells and enhanced the apoptosis ability. Conclusion Limonin can inhibit the malignant biological behavior of NSCLC cells, such as proliferation, migration, and invasion, by inhibiting the JAK2/STAT3 pathway.

Background and objective: Osteosarcoma is a common primary malignant tumor of bone, and doxorubicin is one of the most widely used therapeutic drugs. While the problem of doxorubicin resistance limits the long-term treatment benefits in osteosarcoma patients. The role of miRNAs and their target genes in osteosarcoma have become increasingly prominent. Currently, there is no report on miR-506-3p reversing doxorubicin resistance by targeting STAT3 in osteosarcoma. The purpose of this study was to investigate the molecular mechanism that overexpression of miR-506-3p reverses doxorubicin resistance in drug-resistant osteosarcoma cells. Methods: Doxorubicin-resistant osteosarcoma cells (U-2OS/Dox) were constructed by intermittent stepwise increasing stoichiometry. The target genes of miR-506-3p were predicted by bioinformatics approach and the targeting relationship between miR-506-3p and STAT3 was detected using dual luciferase reporter assay. U-2OS/Dox cells were treated with miR-506-3p overexpression and STAT3 silencing respectively. Then Western blot and RTqPCR were used to detect the protein and mRNA expression levels of JAK2/ STAT3 signaling pathway, drug-resistant and apoptotic associated molecules. The migration and invasion were assessed by cell scratch assay and transwell assay. The cell proliferative viability and apoptosis were investigated by CCK8 assay and flow cytometry assay. Results: U-2OS/Dox cells were successfully constructed with a 14.4-fold resistance. MiR-506-3p is directly bound to the 3′-UTR of STAT3 mRNA. Compared with U-2OS cells, the mRNA expression of miR-506-3p was reduced in U-2OS/Dox cells. Overexpression of miR-506-3p decreased the mRNA expression levels of JAK2, STAT3, MDR1/ABCB1, MRP1/ABCC1, Survivin and Bcl-2, and decreased the protein expression levels of p-JAK2, STAT3, MDR1/ ABCB1, MRP1/ABCC1, Survivin and Bcl-2, and conversely increased Bax expression. It also inhibited the proliferation, migration and invasion of U- 2OS/Dox cells and promoted cells apoptosis. The results of STAT3 silencing experiments in the above indicators were consistent with that of miR-506-3p overexpression. Conclusion: Overexpression of miR-506-3p could inhibit the JAK2/ STAT3 pathway and the malignant biological behaviors, then further reverse doxorubicin resistance in drug-resistant osteosarcoma cells. The study reported a new molecular mechanism for reversing the resistance of osteosarcoma to doxorubicin chemotherapy and provided theoretical support for solving the clinical problems of doxorubicin resistance in osteosarcoma. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective To investigate the influence of limonin on the malignant biological behavior of nonsmall cell lung cancer (NSCLC) cells by regulating the protein tyrosine kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Methods CCK-8 method was applied to detect the survival rate of A549 cells treated with different concentrations of limonin (0, 5, 10, 25, 50, 75, 100 μmol/L). A549 cells were separated into normal culture (NC) group, low-dose limonin group (treatment with 10 µmol/L limonin for 24 h), medium-dose limonin group (treatment with 25 µmol/L limonin for 24 h), high-dose limonin group (treatment with 50 µmol/L limonin for 24 h), coumermycin A1 group (treatment with 10 μmol/L JAK2 activator coumermycin A1+50 μmol/L limonin for 24 h), and AG490 group (treatment with 10 μmol/L JAK2 inhibitor AG490+50 μmol/L limonin for 24 h). Clone formation assay was applied to detect the clones of each group of cells. Transwell assay was applied to detect cell migration and invasion, and flow cytometry was applied to detect apoptosis. Western blot analysis was applied to detect the protein expression levels of JAK2, p-JAK2, STAT3, p-STAT3, E-cadherin, N-cadherin, and vimentin in each group. Results The viability of A549 cells decreased significantly in a limonin concentration-dependent manner (P<0.05), with IC50 of 45.16±1.66 μmol/L. Concentrations of 10, 25, and 50 μmol/L were selected for subsequent experiments. The numbers of clones, migration, and invasion of A549 cells and the protein expression levels of IL-6, p-JAK2, p-STAT3, N-cadherin, and vimentin in the low-, medium-, and high-dose limonin groups significantly decreased, compared with those in the NC group, and the apoptosis rate and E-cadherin protein expression significantly increased (P<0.05). The JAK2 activator coumermycin A1 attenuated the ability of limonin to inhibit the proliferation, migration, invasion, and other malignant biological behavior of A549 cells and attenuated the apoptosis ability. The JAK2 inhibitor AG490 enhanced the ability of limonin to inhibit the proliferation, migration, invasion, and other malignant biological behavior of A549 cells and enhanced the apoptosis ability. Conclusion Limonin can inhibit the malignant biological behavior of NSCLC cells, such as proliferation, migration, and invasion, by inhibiting the JAK2/STAT3 pathway. [ABSTRACT FROM AUTHOR]

Di-(2-ethylhexyl) phthalate (DEHP), a common endocrine-disrupting chemical (EDC), is widely used in daily articles, early exposure to DEHP is associated with many behavioral changes in pups. This study aimed to investigate the effects and underlying mechanisms of maternal exposure to DEHP on the impaired social interaction in pups. Pregnant rats were administered 0, 30, 300, or 750 mg/kg/d DEHP daily by oral gavage. Highly aggressive proliferating immortalized (HAPI) cells were treated with mono-(2-ethylhexyl) phthalate (MEHP) and tyrosine phosphorylation inhibitor (AG490). Our results showed that DEHP exposure induced the activation of microglias (MGs) via activating the janus kinase 2 / signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway, and increased the level of pro-inflammatory factors, then impaired the social behavior in male pups, but not female pups. Moreover, MEHP exposure could also activate HAPI via activating this signaling pathway, and AG490 could inhibit the activation of this signaling pathway caused by MEHP. Therefore, we indicated that maternal exposure to DEHP could cause the gender-specific impaired social interaction in pups that might be related to the activation of MGs.

Autophagy is a vital cellular process that functions to degrade and recycle damaged organelles into basic metabolites. This allows a cell to adapt to a diverse range of challenging conditions. Autophagy assists in maintaining homeostasis, and it is tightly regulated by the cell. The disruption of autophagy has been associated with many diseases, such as neurodegenerative disorders and cancer. This review will center its discussion on providing an in-depth analysis of the current molecular understanding of autophagy and its relevance to brain tumors. We will delve into the current literature regarding the role of autophagy in glioma pathogenesis by exploring the major pathways of JAK2/STAT3 and PI3K/AKT/mTOR and summarizing the current therapeutic interventions and strategies for glioma treatment. These treatments will be evaluated on their potential for autophagy induction and the challenges associated with their utilization. By understanding the mechanism of autophagy, clinical applications for future therapeutics in treating gliomas can be better targeted.

Background: The haungqing (Scutellariae Radix) and baishao (Paeoniae Radix Alba) herb pair (HBHP) is a common prescribed herbal formula or is added to other traditional Chinese medicine (TCM) prescriptions to treat ulcerative colitis (UC). However, the underlying mechanism is unclear. Purpose: Elucidate the efficacy and potential mechanism of HBHP against UC. Methods: First, The UC model of mice induced by dextran sulfate sodium (DSS) was established. The mice were randomly divided into Control group, DSS group, SASP group (390 mg/kg), and HPHP group (1.95 g/kg), with 8 mice per group. Drugs were administrated via oral gavage for 7 days. Then, Disease activity index (DAI), length of the colon, histopathology, and changes in inflammatory cytokines in colonic tissues were analyzed to assess the effect of HBHP on UC. Besides, Network pharmacology was applied to identify the active compounds, core targets of HBHP in the treatment of UC, and the corresponding signaling pathways to explore the underlying mechanisms. Finally, Western blot (WB), immunohistochemistry (IHC) and molecular docking were performed to validate the results. Results: HBHP significantly reduced DAI score and decreased colon length shortening in DSS-induced UC mice. The administration of HBHP was able to effectively alleviated mucosal ulceration and epithelial destruction. In addition, HBHP treatment obviously - reduced the expressions of TNF-α, IL-6, and IL-1β in colon tissues (p

AIM: To investigate the effect of kinesin family member 20A（ KIF20A） on the malignant behavior and chemoresistance of colorectal cancer （CRC）, and to explore the underlying mechanism. METHODS: Twelve cases of CRC and normal colorectal tissues were obtained from Department of Pathology, Nanfang Hospital, Southern Medical University. The KIF20A expression in CRC tissues and adjacent tissues was detected by immunohistochemistry, Western blot and RT-qPCR. Several CRC cell lines were transfected with KIF20A short hairpin RNA（ sh-KIF20A） or infected with KIF20A overexpression lentivirus （LV-KIF20A）. The RKO or LOVO cells were divided into MOCK group （non-transfected RKO or LOVO cells）, LV-NC group （negative control lentivirus-infected RKO or LOVO cells）, and LV-KIF20A group （LV-KIF20A-infected RKO or LOVO cells）. The HCT-116 cells were divided into MOCK group （non-transfected HCT-116 cells）, sh-NC group （negative control shRNA-transfected HCT-116 cells）, and sh-KIF20A group （sh-KIF20A-transfected HCT-116 cells）. Cell viability and colony formation ability were detected by CCK-8 and colony formation assays. Flow cytometry was used to assess cell apoptosis. The levels of apoptosis-related proteins, including cleaved caspase-3, Bax and Bcl-2, and JAK2/STAT3 signaling pathway-related proteins were detected by Western blot. Immunohistochemistry was used to assess the levels of KIF20A and p-STAT3 in chemotherapy-sensitive and -insensitive CRC tissues. The RKO cells infected with LV-NC or LV-KIF20A were subcutaneously inoculated into the right body of BALB/c nude mice （5 mice per group）, and the effect of KIF20A on tumor growth in vivo was determined. RESULTS: The KIF20A expression was significantly increased in CRC tissues and cells （P<0. 05）. Knockdown of KIF20A significantly inhibited CRC cell viability and colony formation, while KIF20A overexpression significantly increased CRC cell viability and colony formation. With 5-fluorouracil treatment, KIF20A knockdown increased the expression of cleaved caspase-3 and Bax, decreased the expression of Bcl-2, and induced apoptosis. In contrast, KIF20A overexpression inhibited cleaved caspase-3 and Bax expression, increased Bcl-2 expression, and inhibited apoptosis. Overexpression of KIF20A up-regulated JAK2 and STAT3 phosphorylation, while KIF20A knockdown down-regulated JAK2 and STAT3 phosphorylation. Knockdown of STAT3 reversed the increase in proliferation and the inhibition of apoptosis induced by KIF20A overexpression. Furthermore, the levels of KIF20A and p-STAT3 in chemotherapy-insensitive CRC tissues were higher than those in chemotherapy-sensitive CRC tissues. In vivo, KIF20A overexpression promoted CRC tumor growth （P<0. 05）. CONCLUSION: KIF20A regulated malignant behavior and chemoresistance in CRC by activating the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

The aim of this study was to discover new biomarkers to detect breast cancer (BC), which is an aggressive cancer with a high mortality rate. In this study, bioinformatic analyses (differential analysis, weighted gene co-expression network analysis, and machine learning) were performed to identify potential candidate genes for BC to study their molecular mechanisms. Furthermore, Quantitative Real-time PCR and immunohistochemistry assays were used to examine the protein and mRNA expression levels of a particular candidate gene (DLGAP5). And the effects of DLGAP5 on cell proliferation, migration, invasion, and cell cycle were further assessed using the Cell Counting Kit-8 assay, colony formation, Transwell, wound healing, and flow cytometry assays. Moreover, the changes in the JAK2/STAT3 signaling-pathway-related proteins were detected by Western Blot. A total of 44 overlapping genes were obtained by differential analysis and weighted gene co-expression network analysis, of which 25 genes were found in the most tightly connected cluster. Finally, NEK2, CKS2, UHRF1, DLGAP5, and FAM83D were considered as potential biomarkers of BC. Moreover, DLGAP5 was highly expressed in BC. The down-regulation of DLGAP5 may inhibit the proliferation, migration, invasion, and cell cycle of BC cells, and the opposite was true for DLGAP5 overexpression. Correspondingly, silencing or overexpression of the DLGAP5 gene inhibited or activated the JAK2/STAT3 signaling pathway, respectively. DLGAP5, as a potential biomarker of BC, may impact the cell proliferation, migration, invasion, cell cycle, and BC development by modulating the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

Background: Cholangiocarcinoma (CCA) is a malignant epithelial tumor characterized by a dismal prognosis. Given the lack of therapeutic strategies and durable treatment options currently available, identifying innovative treatments for CCA is an urgent unmet clinical need. Cucurbitacin C (CuC) is a distinct variant of the cucurbitacin family, displaying promising anti-cancer activity against various tumor types. The primary objective of our research is to elucidate the promising effects of CuC on CCA. Methods: The impact of CuC on CCA cell lines was assessed by cell count kit-8 assay, EdU staining assay, colony formation assay, wound-healing assay, and Transwell assay. Flow cytometric analysis was conducted to explore the function of CuC treatments on cell-cycle distribution and apoptosis in CCA cells. Computational biology and network pharmacology approaches were utilized to predict potential targets of CuC. Furthermore, a tumor xenograft mouse model was established using CCA cells to explore the anti-cancer effects of CuC in vivo. Results: Our research findings revealed that CuC exerted a suppressive effect on CCA cell progression. Cell viability assays, EdU staining assays, and colony formation assays demonstrated that CuC effectively suppressed viability and proliferation of CCA cells. Wound-healing assays and Transwell assays indicated that CuC effectively inhibits the migratory and invasive capabilities of CCA cells. Flow cytometry analysis elucidated that CuC played its anti-proliferative role in CCA cells by arresting G0/G1 phase and increasing apoptosis. Through bioinformatics and network pharmacology analysis, in conjunction with western blot analysis, we demonstrated CuC mediated the inhibition of CCA cell progression through modulation of JAK2/STAT3 pathway. Additionally, the CCA xenograft tumor model was established, and the results supported the inhibition of CuC treatment against CCA progression in vivo. Conclusion: Our study demonstrates that CuC possesses notable capabilities to suppress cell proliferation, migration, and invasion in CCA. Importantly, the inhibitory effects of CuC on CCA progression are attributed to its modulation of the JAK2/STAT3 signaling pathway. Altogether, our study demonstrated that CuC holds promise as a prospective therapeutic agent for treating CCA. [ABSTRACT FROM AUTHOR]

Objective To investigate the regulatory effect of JAK2/STAT3 signaling pathway inhibitor AG490 on the functional phenotype of M2-like macrophages and its effects on proliferation, apoptosis, migration, and invasion of gastric cancer cells. Methods Human mononuclear cell line THP-1 was induced to differentiate into M0 macrophages by PMA in vitro. The M1-like phenotype was induced by LPS and IFN-γ, and M2-like phenotype was induced by IL-4 and IL-13, respectively, and identified by immunofluorescence labeling CD68, CD86 and CD206. The mRNA expressions of CD163, Arg1, CCL22, PPARγ, IL-10, IL-20 and TNF-α were determined by RT-qPCR. The expressions of key proteins in JAK2/STAT3 signaling pathway were detected by Western blotting. M2-like macrophages were treated with JAK2/STAT3 inhibitor(AG490)to observe the expression level of marker genes for M2 like phenotype. Macrophages were co-cultured with gastric cancer cells, and the effects of the macrophages on proliferation, migration, invasion, and apoptosis of gastric cancer cells were detected by CCK-8 method, healing assay, transwell intracellular Matrigel invasion assay, and flow cytometry. The xenograft tumor model of MKN45 gastric cancer in nude mice was prepared, and the tumor size and quality were observed for 20 days after the model was established. Results THP-1 cells were induced into M1-like macrophages and M2-like macrophages. M1-like marker(CD86)and M2-like marker(CD206)were identified by flow cytometry. The P38MAPK, JAK2, p-STAT3/STAT3 protein levels of M2-like macrophages treated with AG490 were significantly reduced. The mRNA expression levels of Arg1, CCL22, PPARγ and IL-10 were significantly reduced in the group of M2-like macrophages treated with AG490. Co-culture of M2-like macrophages with gastric cancer cells could promote gastric cancer cell viability, increase migration and invasion ability, and inhibit apoptosis. When the group of M2-like macrophages treated with AG490 was co-cultured with gastric cancer cells, the proliferation activity of MKN-45 cells and MGC823 was significantly lower than that in M2 group(1.047±0.062 vs. 1.426±0.076, 1.149±0.006 vs. 1.301±0.015). Compared to M2 group, the migration(100.0%±5.73% vs. 72%±3.85%)and invasion ability(100.0%±7.40% vs. 60%±6.54%)of MGC823 gastric cancer cells in AG490 treatment group were significantly reduced. The apoptosis rate of MGC823 cells in the AG490 treated group was significantly higher than that in M2 group(27.51%±0.70% vs. 20.82%±0.92%). In the nude mouse xenograft tumor model, the volume and weight of the transplanted tumor collected at day 20 were significantly lower in AG490 treated group than in M2 group(736.04±182.34 vs. 1 080.5±250.57)mm³, (0.64±0.11 vs. 0.87±0.17)g. Conclusion AG490 downregulates the activation level of JAK2/STAT3 signaling pathway in M2-like macrophages, inhibits M2-type polarization, partially reverses the cancer-promoting function of M2-like macrophages, inhibits proliferation, migration and invasion of gastric cancer cells, and induces apoptosis. The JAK2/STAT3 signaling pathway can be further studied as a potential therapeutic target for gastric cancer. [ABSTRACT FROM AUTHOR]

Acacetin is a natural flavonoid compound found in diverse plants, which has strong anti‐inflammatory and anti‐cancer activities. This work aimed at investigating how acacetin functions on esophageal squamous carcinoma cells. In this work, esophageal squamous carcinoma cell lines were subjected to increasing doses of acacetin, and the proliferative, migrative, invasive and apoptotic phenotypes were evaluated by a series of in vitro experiments. Genes related to acacetin and esophageal cancer were predicted by bioinformatics analysis. The levels of apoptosis‐relevant proteins and JAK2/STAT3 pathway‐relevant proteins in esophageal squamous carcinoma cells were probed by Western blot. It was revealed that acacetin could block the growth and aggressiveness of TE‐1 and TE‐10 cells and promote the apoptosis. Acacetin treatment induced bax's expression and repressed bcl‐2's expression. Notably, acacetin inhibits JAK2/STAT3 pathway in esophageal squamous carcinoma cells. In summary, acacetin inhibits the malignant progression of esophageal squamous carcinoma via restraining JAK2/STAT3 signaling. [ABSTRACT FROM AUTHOR]

Diabetic cardiomyopathy (DCM) represents a common pathological state brought about by diabetes mellitus (DM). Patchouli alcohol (PatA) is known for its diverse advantageous effects, notably its anti-inflammatory properties and protective role against metabolic disorders. Despite this, the influence of PatA on DCM remains relatively unexplored. To explore the effect of PatA on diabetes-induced cardiac injury and dysfunction in mice, streptozotocin (STZ) was used to mimic type 1 diabetes in mice. Serological markers and echocardiography show that PatA treatment protects the heart against cardiomyopathy by controlling myocardial fibrosis but not by reducing hyperglycemia in diabetic mice. Discovery Studio 2017 software was used to perform reverse target screening of PatA, and we found that JAK2 may be a potential target of PatA. RNA-seq analysis of heart tissues revealed that PatA activity in the myocardium was primarily associated with the inflammatory fibrosis through the Janus tyrosine kinase 2 (JAK2)/signal transducer and activator of the transcription 3 (STAT3) pathway. In vitro, we also found that PatA alleviates high glucose (HG) + palmitic acid (PA)-induced fibrotic and inflammatory responses via inhibiting the JAK2/STAT3 signaling pathway in H9C2 cells. Our findings illustrate that PatA mitigates the effects of HG + PA- or STZ-induced cardiomyopathy by acting on the JAK2/STAT3 signaling pathway. These insights indicate that PatA could potentially serve as a therapeutic agent for DCM treatment.

Suppressor of cytokine signaling 3 (SOCS3) is a negative regulatory protein that has been identified as a key inhibitory regulator of JAK/STAT signaling pathway. However, the mutual regulatory relationship between SOCS3 and JAK2/STAT3 signaling pathway after vocal fold injury remains unclear. In this study, we used small interfering RNA (siRNA) to investigate the mechanism of SOCS3 regulating of fibroblasts through JAK2/STAT3 signaling pathway after vocal fold injury. Our data shows that SOCS3 silencing promotes the transformation of normal vocal fold fibroblasts (VFFs) into an fibrotic phenotype and activates the JAK2/STAT3 signaling pathway. JAK2 silencing significantly inhibits the increase in type I collagen and α-smooth muscle actin (α-SMA) secretion in VFFs induced by TGF-β but has no significant effect on normal VFFs. The silencing of SOCS3 and JAK2 reverses the fibrotic phenotype of VFFs induced by SOCS3 silencing. Therefore, we suggest that SOCS3 can affect the activation of vocal fold fibroblasts by regulating the JAK2/STAT3 signaling pathway after vocal fold injury. It provides a new insight for promoting the repair of vocal fold injury and preventing the formation of fibrosis. [ABSTRACT FROM AUTHOR]

The highly conserved ribosomal protein L34 (RPL34) has been reported to play an essential role in the progression of diverse malignancies. RPL34 is aberrantly expressed in multiple cancers, although its significant in colorectal cancer (CRC) is currently unclear. Here, we demonstrated that RPL34 expression was higher in CRC tissues than in normal tissues. Upon RPL34 overexpression, the ability of proliferation, migration, invasion, and metastasis of CRC cells were significantly enhanced in vitro and in vivo. Furthermore, high expression of RPL34 accelerated cell cycle progression, activated the JAK2/STAT3 signaling pathway, and induced the epithelial-to-mesenchymal transition (EMT) program. Conversely, RPL34 silencing inhibited the CRC malignant progression. Utilizing immunoprecipitation assays, we identified the RPL34 interactor, the cullin-associated NEDD8-dissociated protein 1 (CAND1), which is a negative regulator of cullin-RING ligases. CAND1 overexpression reduced the ubiquitin level of RPL34 and stabilized RPL34 protein. CAND1 silencing in CRC cells resulted in a decrease in the ability of proliferation, migration, and invasion. CAND1 overexpression promoted CRC malignant phenotypes and induced EMT, and RPL34 knockdown rescued CAND1-induced CRC progression. In summary, our study indicates that RPL34 acts as a mediator, is stabilized by CAND1, and promotes proliferation and metastasis, in part, through the activation of the JAK2/STAT3 signaling pathway and induction of EMT in CRC. [ABSTRACT FROM AUTHOR]

To investigate the effects of quercetin on human chondrosarcoma cell migration, invasion and epithelial intersti- tial transformation (EMT) and the regulatory role of Janus kinase 2/signal transduction and transcription promoter 3 (JAK2/ STAT3) signaling pathway, human chondrosarcoma SW-1353 cells were cultured in vitro and treated with quercetin of differ- ent concentrations for 24 h. The optimal concentration of quercetin (50.0 μmol/L) was selected by cell counting kit (CCK-8) for subsequent experiments. The cells were divided into control group with no intervention, quercetin group with 50.0 μmol/L quercetin, positive control group with 6.0 μmol/L adriamycin, inhibitor group involving 50.0 μmol/L quercetin +50.0 μmol/L JAK2/STAT3 pathway inhibitor AG490, and activator group involving 50.0 μmol/L quercetin +0.5 μmol/L JAK2/STAT3 path- way activator colivelin. After 24 h of intervention, cell morphology, migration, invasion, and expression levels of JAK2/STAT3 pathway and EMT-related proteins were analyzed by inverted microscope, Transwell chamber and Western blot. The experimental results showed that the optimum concentration of quercetin was determined by CCK-8 method to be 50.0 μmol/L. Compared with the control group, the growth of quercetin group were positive control group were inhibited, the cell migration number, invasion number, expression levels of N-cadherin, Vimentin, fibronectin (FN), p-JAK2 and p-STAT3 in quercetin group and positive control group decreased (P<0.05), E-cadherin expression level increased (P<0.05); compared with quercetin group, the change trend of the above indexes in inhibitor group was more significant (P<0.05), while the change of these indexes in activator group was reversed (P<0.05). Quercetin can inhibit the growth, migration, invasion and EMT progression of human chondrosarcoma SW-1353 cells, possibly by blocking the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

Cadmium (Cd), a common heavy metal in the environment, is associated with cognitive impairment. In the present study, we carried out a preliminary inquiry to explore whether Cd causes neurotoxicity by regulating the JAK2/STAT3 signaling pathway and affecting the expression of klotho genes in vivo and in vitro, providing clues for the mechanism of Cd-induced cognitive dysfunction. The rat samples were injected with Cd chloride solution for 14 weeks, and the memory function of the rats was detected. Different concentrations of Cd and JAK2/STAT3 signaling pathway inhibitors were used to treat PC12 cells and thus detect the apoptosis rate. The protein expression levels of JAK2, p-JAK2, STAT3, p-STAT3, and klotho in rat and PC12 cell were detected by ELISA and Western blot, respectively. With the increase in exposure dose, the memory function of rats was severely impaired. The expression of p-JAK2 and p-STAT3 proteins was significantly up-regulated, whereas that of klotho was significantly down-regulated both in vivo and in vitro (p < 0.05). In comparison with the high-dose Cd exposure group, after adding tyrphostin AG490 (AG490), the apoptosis rate of PC12 cells increased, whereas the phosphorylation levels of JAK2 and STAT3 in the cells decreased significantly (p < 0.05). Cd exposure may cause neurotoxicity by regulating the JAK2/STAT3 signaling pathway and down-regulating klotho protein expression, leading to cognitive dysfunction. [ABSTRACT FROM AUTHOR]

Depression is a serious disorder with a large impact on both an individual's quality of life and society as a whole. This study aimed to evaluate the potential involvement of nuclear factor kappa-B (NF-kB) and the Janus activated kinase (JAK) and Signal transducer and activator of transcription proteins (STAT) signaling pathway in the pathogenesis of genetically predisposed depression in female rats. The obtained results showed increased phosphorylation in JAK2 and STAT3, and increased protein levels of NF-kB in the hippocampus of Wistar Kyoto rats compared to Wistar rats. These results suggest that disturbance in these pathways could have a significant role in the pathophysiology of genetically predisposed depression in females.

Objective To investigate the mechanism of Elabela in myocardial ischemia / reperfusion injury. Methods The concentrations of Elabela in patients with acute ST-segment elevation myocardial infarction （STEMI） were detected. The expression and function of Elabela in H9C2 cells （rat embryonic heart clone cell line） were evaluated by obtaining clinical blood samples and establishing an oxidative stress model in vitro to simulate ischemia / reperfusion injury. Results The concentration of Elabela in the plasma of STEMI patients and the expression of Elabela in H9C2 cells treated with H2O2 were both up-regulated （both P < 0.001）. Elabela could reduce the level of Bax, the rates of cardiomyocyte apoptosis, the number of reactive oxygen species-positive cells, the levels of lactate dehydrogenase and malondialdehyde, whereas increased superoxide dismutase activity and Bcl-2 expression in H2O2 treated H9C2 cells （all P < 0.05）. The inhibition of α7nAChR/JAK2/STAT3 signaling pathway by the specific inhibitor α-ring snake toxin partially counteracted the protective effect of Elablela. Conclusion Elabela ameliorates H2O2-induced H9C2 cell injury by activating the α7nAChR/JAK2/STAT3 signaling pathway.

AIM: To explore the effect of Yizheng Prescription on immune function in rats with repeated respiratory tract infection （RRTI） of lung-spleen Qi deficiency type and its molecular mechanism. METHODS: Sixty SD rats were divided into control group, RRTI group, low-, medium- and high-dose Yizheng Prescription groups （intragastric gavage of 10, 20 and 30 g/kg Yizheng Prescription crude drug）, and Janus kinase 2 （JAK2） inhibitor AG490 group （intraperitoneal injection of 5 mg/kg AG490）, with 10 rats in each group. Except for the control group, lung-spleen Qi deficiency type RRTI model was established in the rats of the other groups using fatigue, diet disorder and wood shaving+tobacco smoke method. The general state of the rats was observed. The lung function ［forced expiratory volume in one second （FEV1）, forced vital capacity（ FVC） and peak expiratory flow rate（ PEF）］ was measured. The spleen and thymus indexes were calculated. The pathological changes of lung tissues were observed by HE staining. The levels of immunoglobulin A （IgA）, IgM, IgG, tumor necrosis factor-α（ TNF-α）, interleukin-4（ IL-4） and IL-17 in serum, and secretory IgA（ sIgA） in bronchoalveolar lavage fluid （BALF） were detected by ELISA. The percentages of CD3+, CD4+ and CD8+ T-lymphocytes in peripheral blood were detected by flow cytometry. The expression levels of JAK2/signal transducer and activator of transcription 3 （STAT3） signaling pathway-related proteins in lung tissues were detected by Western blot. RESULTS: Compared with control group, the rats in RRTI group had poorer general status and obvious pathological damage in the lung tissue. Meanwhile, the lung injury score, the FEV1, FVC, FEV1/FVC and PEF levels, spleen and thymus indexes, serum IgA, IgM, IgG and IL-4 levels, BALF sIgA levels, and peripheral blood CD3+, CD4+, CD8+ and CD4+/CD8+ T-lymphocytes in RRTI group were significantly decreased, while serum TNF-α and IL-17 levels, and ratios of p-JAK2/JAK2 and p-STAT3/STAT3 in lung tissue were significantly increased （P<0. 05）. Compared with RRTI group, the general state of the rats in low-, medium- and high-dose Yizheng Prescription groups and JAK2 inhibitor AG490 group was improved, and the pathological damage of lung tissue was attenuated to varying degrees. Meanwhile, the lung injury score, the FEV1, FVC, FEV1/FVC and PEF levels, spleen and thymus indexes, serum IgA, IgM, IgG and IL-4 levels, BALF sIgA levels, and peripheral blood CD3+, CD4+, CD8+ and CD4+/CD8+ T-lymphocytes were significantly increased, while serum TNF-α and IL-17 levels and ratios of p-JAK2/JAK2 and p-STAT3/STAT3 in lung tissue were significantly decreased （P< 0. 05）. The effects in Yizheng Prescription groups were dose-dependent. CONCLUSION: Yizheng Prescription alleviates lung injury and improves immune function in RRTI rats of lung-spleen Qi deficiency type. Its mechanism may be related to inhibiting the activation of JAK2/STAT3 signaling pathway, and reducing systemic inflammatory response. [ABSTRACT FROM AUTHOR]

BACKGROUND: Numerous studies have shown that mesenchymal stem cells can treat autoimmune diseases through a variety of immunomodulatory pathways, and genetic modification may improve the therapeutic potential of mesenchymal stem cells. MiR-1-5p is lowly expressed in systemic lupus erythematosus, and mesenchymal stem cells overexpressing miR-1-5p may have a therapeutic effect on systemic lupus erythematosus. OBJECTIVE: To investigate whether miR-1-5p-modified umbilical cord mesenchymal stem cells mediate the JAK2/STAT3 pathway and have a therapeutic effect on systemic lupus erythematosus MRL/lpr model mice. METHODS: Twenty MRL/lpr lupus mice were randomly divided into miR-1-5p transfected umbilical cord mesenchymal stem cells treatment group (UCMSCs+miR-1-5p group), miRNA negative control transfected umbilical cord mesenchymal stem cells treatment group (UC-MSCs+miR-NC group), umbilical cord mesenchymal stem cell treatment group (UC-MSCs group), PBS treatment group (PBS group), and MRL/lpr lupus mice untreated (control group). MRL/lpr lupus mice were treated with tail vein transplantation, once a week, 5 times. Mice were sacrificed at 1-week intervals after the end of treatment. Hematoxylin-eosin staining was used to observe the pathological changes of liver tissue and intestinal tissue of MRL/lpr mice in each group. Western blot assay was used to detect JAK2/STAT3 pathway proteins and their phosphorylation levels and expression levels of interleukin-18 in kidney tissue of mice in each group. RESULTS AND CONCLUSION: (1) Compared with control group, the infiltration of inflammatory cells in liver tissue of MRL/lpr mice in UC-MSCs+miR-1-5p group, UC-MSCs+miR-NC group, and UC-MSCs group decreased significantly, and hepatocyte edema and degeneration could be seen in UC-MSCs+miR-NC group. Goblet cells in intestinal tissue of MRL/lpr mice in UC-MSCs+miR-1-5p group increased significantly, but there was no significant difference among other groups. (2) Compared with control group, the expression of p-JAK2 (Y1007+Y1008), p-STAT3 (s727) and interleukin-18 in renal tissue of lupus mice in UC-MSCs+miR1-5p group was significantly down-regulated (P < 0.05); p-STAT3(s727) (P < 0.05) and interleukin-18 (P < 0.01) were significantly down-regulated in the kidney tissue of lupus mice in UC-MSCs group; expression level of interleukin-18 in UC-MSCs+miR-NC group was significantly down-regulated (P < 0.05); and there was no significant difference in the expression of p-STAT3 (Y705) in renal tissue of lupus mice of each group (P > 0.05). The expression of interleukin-18 in UC-MSCs group was significantly lower than that in PBS group (P < 0.05). (3) The results suggested that umbilical cord mesenchymal stem cells modified by miR-1-5p may alleviate liver and intestinal damage in MRL/lpr mice by inhibiting the activation of JAK2/STAT3 pathway and the expression of interleukin 18, thereby playing a certain therapeutic role in lupus mice. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of LncRNA-NEAT1 on JAK2/STAT3 signaling pathway, inflammatory response and pregnancy outcome in hypertensive rats during pregnancy. Methods: The hypertensive rat model of pregnancy was established by injecting L-arginine methyl ester. The protein expression of JAK2/STAT3 signaling pathway was detected by Western blot. Inflammatory factors and vascular endothelial injury factors were detected by ELISA. Proteinuria, caudal venous pressure and stillbirth rate were observed and recorded. Results: Compared with blank group, the protein expression levels of JAK2 and STAT3 in model group, LncRNA-NEAT1 overexpression group and LncRNA-NEAT1 inhibition group were significantly higher (P＜0.05). Compared with model group, the protein expression levels of JAK2 and STAT3 in LncRNA-NEAT1 inhibition group were significantly lower (P＜0.05), while the protein expression levels of JAK2 and STAT3 in LncRNA-NEAT1 overexpression group were significantly higher (P＜0.05). Compared with blank group, the levels of ET-1 and s ICAM-1 in model group, LncRNA-NEAT1 overexpression group and LncRNA-NEAT1 inhibition group were significantly higher, while the level of NO was significantly lower (P＜0.05). Compared with model group, the levels of ET-1 and s ICAM-1 in LncRNA-NEAT1 overexpression group and LncRNA-NEAT1 inhibition group were significantly higher (P＜0.05), while the level of NO was significantly lower (P＜0.05). The expression levels of ET-1 and s ICAM-1 in LncRNA-NEAT1 overexpression group were significantly higher, but NO was significantly lower (P＜0.05). Compared with blank group,the expression levels of TNF-α, IL-1β and IL-18 in model group, LncRNA-NEAT1 overexpression group and LncRNA-NEAT1inhibition group were significantly higher (P＜0.05). Compared with model group, the expression levels of TNF-α, IL-1β and IL-18 in LncRNA-NEAT1 inhibition group were significantly lower (P＜0.05), while the expression levels of TNF-α, IL-1β and IL-18 in LncRNA-NEAT1 overexpression group were significantly higher (P＜0.05). Compared with blank group, caudal venous pressure and 24 h proteinuria levels in model group, LncRNA-NEAT1 overexpression group and LncRNA-NEAT1 inhibition group were significantly higher (P＜0.05). Compared with model group, the caudal venous pressure and 24 h proteinuria expression levels in LncRNA-NEAT1inhibited group were significantly lower (P＜0.05), while the caudal venous pressure and 24 h proteinuria expression levels in LncRNA-NEAT1 overexpression group were significantly higher (P＜0.05). The stillbirth rate of LncRNA-NEAT1 overexpression group (21.56%) was higher than that of model group (16.72%) and LncRNA-NEAT1 inhibition group (5.65%). Conclusions: Down-regulation of LncRNA-NEAT1 in gestational diabetes rats can inhibit the expression of JAK2/STAT3 signaling pathway and down-regulate the expression of downstream pro-inflammatory factors, so as to alleviate vascular endothelial injury and reduce the stillbirth rate. [ABSTRACT FROM AUTHOR]

Lentinan (LNT), an immunoregulatory agent, is a compound derived from the mushroom Lentinus edodes. However, the immunological function and antitumor effect of LNT and the underlying mechanism remain poorly understood. In this study, we aimed to investigate the potential mechanism of LNT in treating TNBC lung metastasis by regulating macrophages in vivo and in vitro. In the Luc-4T1 mouse model, LNT inhibited the metastasis of the triple-negative breast cancer (TNBC) cells into the lungs. Additionally, as demonstrated by the immunohistochemical results and down-regulation of the mRNA expression of M2-type macrophage marker genes, LNT substantially inhibited IL-4-induced M2 macrophage polarization. Molecular docking and western blotting data further suggested that LNT is likely to target activation of the JAK2/STAT3 signaling pathway, and siRNA transfection experiments confirmed these findings. These results provide important implications for the use of LNT as a functional food to enhance immune function in the treatment of breast cancer.

Abstract Background Prostate cancer (PCa) is one of the most common malignant tumors in the male urinary system. In recent years, the morbidity and mortality of PCa have been increasing due to the limited effects of existing treatment strategies. Long non-coding RNA (lncRNA) LINC00893 was reported to inhibit the proliferation and metastasis of papillary thyroid cancer cells, but its role in PCa has not been reported. This study aims to investigate the role and underlying mechanism of LINC00893 in regulating the progression of PCa cells. Methods We first compared LINC00893 expression levels between PCa tissues and normal prostate tissues through TCGA database. The relative LINC00893 expression levels were further validated in 66 pairs of PCa tissues and para-cancerous normal tissues, as well as in PCa cell lines. Gain-of-function experiment was performed by transfecting PCa cell with LINC00893 expression vector, and CCK (Cell count kit)-8, 5-Ethynyl-2′-deoxyuridine (EdU) incorporation, colony information and transwell assays were conducted to assess the functional phenotypes. Dual-luciferase reporter, RNA-binding protein immunoprecipitation (RIP) and RNA pull-down assays were performed to evaluate the molecular interactions. Results LINC00893 was downregulated in PCa tissues and cell lines, and patients with low expression of LINC00893 were associated with a poorer overall survival rate. LINC00893 overexpression hindered the proliferation, epithelial-mesenchymal transition (EMT) as well as the migratory ability of PCa cells, and suppressed the tumorigenesis of PCa cells in nude mice. We further demonstrated that LINC00893 acted as a sponge for miR-3173-5p and inhibited its activity, which in turn regulated the suppressor of cytokine signaling 3 (SOCS3)/Janus Kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling axis. Conclusions Our study demonstrated that LINC00893 suppresses the progression of PCa cells through targeting miR-3173-5p/SOCS3/JAK2/STAT3 axis. Our data uncovers a novel tumor-suppressor role of LINC00893 in PCa, which may serve as a potential strategy for targeted therapy in PCa. Grapical Abstract

Inter-α-trypsin inhibitor heavy chain 4 (ITI-H4) is one of the acute phase proteins and is mainly related with inflammatory diseases such as bacterial bloodstream infection and recurrent pregnancy loss (RPL). In a previous study, ITI-H4 was reported to be cleaved by kallikrein B1 (KLKB1) and its cleaved form induces the imbalance between pro- and anti-inflammatory cytokines. Therefore, in this study, putative substrates of ITI-H4 were isolated by immunoprecipitation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) analysis. Of those, phosphoglycerate kinase 1 (PGK1) was found to be a binding protein of ITI-H4. PGK1 increases the level of ITI-H4 expression and blocks the cleavage of ITI-H4 mediated by KLKB1. It also inhibits pro-inflammatory response by inhibiting the JAK2/STAT3 signaling pathway. Therefore, PGK1, a novel binding partner of ITI-H4, is expected to have cellular functions in the pathogenesis of ITI-H4-related inflammatory diseases.

Bee pollen possesses an anti-cardiomyocyte injury effect by reducing oxidative stress levels and inhibiting inflammatory response and apoptosis, but the possible effect mechanism has rarely been reported. This paper explores the effect of the extract of lotus bee pollen (LBPE) on cardiomyocyte hypertrophy (CH) and its mechanism. The main components of LBPE were identified via UPLC-QTOF MS. An isoproterenol-induced rat H9c2 CH model was subsequently used to evaluate the protection of LBPE on cells. LBPE (100, 250 and 500 μg∙mL−1) reduced the surface area, total protein content and MDA content, and increased SOD activity and GSH content in CH model in a dose-dependent manner. Meanwhile, quantitative real-time PCR trials confirmed that LBPE reduced the gene expression levels of CH markers, pro-inflammatory cytokines and pro-apoptosis factors, and increased the Bcl-2 mRNA expression and Bcl-2/Bax ratio in a dose-dependent manner. Furthermore, target fishing, bioinformatics analysis and molecular docking suggested JAK2 could be a pivotal target protein for the main active ingredients in the LBPE against CH. Ultimately, Western blot (WB) trials confirmed that LBPE can dose-dependently inhibit the phosphorylation of JAK2 and STAT3. The results show that LBPE can protect against ISO-induced CH, possibly via targeting the JAK2/STAT3 pathway, also suggesting that LBPE may be a promising candidate against CH. [ABSTRACT FROM AUTHOR]

Objective: To study the effect of plumbagin (PL) on the migration and invasion of human hepatocellular carcinoma (HCC) cells and its possible mechanism. Methods: The cell counting kit (CCK-8) was used to detect the effects of different concentrations of plumbagin on the proliferation of human hepatocellular carcinoma Huh-7 and LM3 cells. The effect of plumbagin on the migration ability of Huh-7 and LM3 cells was detected by scratch test and Transwell migration test, and the effect of on the invasion ability of Huh-7 and LM3 cells was detected by Transwell invasion test. Western Blot was used to detect the expression of E-cadherin, N-cadherin, matrix metalloproteinase-2 and related proteins in JAK2/STAT3 signaling pathway in Huh-7 and LM3 cells. Results: Plumbagin could inhibit the proliferation of Huh-7 and LM3 cells in a time-and concentration-dependent manner. Plumbagin inhibited the migration and invasion of Huh-7 and LM3 cells in a concentration dependent manner, and it can down-regulate the expression of N-cadherin and MMP-2 protein, up-regulate the expression of E-cadherin protein, and inhibit the activation of JAK2/STAT3 signaling pathway. Conclusion: Plumbagin can inhibit the migration and invasion of human hepatocellular carcinoma Huh-7 and LM3 cells, and the molecular mechanism of this process may be related to the inhibition of JAK2/STAT3 signaling pathway activation. [ABSTRACT FROM AUTHOR]

Objective To explore the role of elevated serum interleukin 1(IL-1) levels in patients with gastric cancer and activation of JAK2/STAT3 signaling pathway in tissues of gastric cancer. Methods A total of 30 gastric cancer surgery patients and 30 healthy subjects were selected from Wuwei Cancer Hospital of Gansu Province; The levels of IL-1 and IL-10 in serum was determined by enzyme-linked immunosorbent assay (ELISA); Immunohistochemistry and Western blot were used to detect the contents of Bcl-2, Bax and JAK2,STAT3, pJAK2, pSTAT3, Bcl-2, Bax in tissues. The contents of JAK2, STAT3, Bcl-2 and Bax genes were measured by RT-qPCR. Results Compared with the control group, the level of IL-1 in the serum of patients with gastric cancer was up-regulated, and the level of IL-10 was down-regulated(P＜0.05); The content of Bcl-2 in cancer tissue was significantly higher than that in the normal group and the adjacent group, but the Bax content showed an opposite profile(P＜0.05). Compared with the control group, JAK2, STAT3, pJAK2, pSTAT3 and Bcl-2 protein content in cancer tissue and adjacent tissues were increased, while the content of Bax was decreased(P＜0.05). Compared with the adjacent tissues, the JAK2, STAT3, pJAK2, pSTAT3 and Bcl-2 protein contents in cancer tissues increased, while the Bax content was decreased(P＜0.05). Compared with the control group, the contents of JAK2, STAT3 and Bcl-2 mRNA in cancer tissues and adjacent tissues were increased, while Bax mRNA was decreased(P＜0.05). Compared with adjacent tissues, the contents of JAK2, STAT3 and Bcl-2 mRNA in cancer tissues increased, while the content of Bax mRNA was decreased(P＜0.05). Conclusions Abnormal elevation of proinflammatory factors and abnormal activation of JAK2/STAT3 signaling pathway are closely related to the occurrence and development of gastric cancer.

Cognitive impairment (CI) is now well-accepted as a complication and comorbidity of diabetes mellitus (DM), becoming a serious medical and social problem. Jiao-tai-wan (JTW), one of noted traditional Chinese medicine (TCM), showed dual therapeutic effects on DM and CI. Nevertheless, the potential mechanism is unclear. This study sought to investigate the mechanism how JTW protected against DM and CI and screen the active component in JTW. Db/db mice were used as mouse models. Mice were treated by gavage with 0.9 % saline (0.1 mL/10g/d), low dose of JTW (2.4 g/kg/d) or high dose of JTW (4.8 g/kg/d) for 8 weeks separately. To access the effects of JTW, the levels of OGTT, HOMA-IR, blood lipids, inflammatory cytokines in serum and hippocampus were measured, behavioral tests were conducted, and histopathological changes were observed. The mechanism exploration was performed via network pharmacology, RT-qPCR, western blot, and immunofluorescence staining (IF). The impact and mechanism of coptisine in vitro were investigated using BV2 cells induced by LPS as cellular models. In vitro experiments were conducted in two parts. The first part comprised four groups: Control group, LPS group, LPS+LCOP group and LPS+HCOP group. The second part consisted of four groups: Control group, LPS group, LPS+HCOP group, and LPS+ Fed group. The western blot and RT-qPCR methods were used to examine the changes in biomarkers of the JAK2/STAT3 signaling pathways in BV2 cells. The results demonstrated that JTW could improve OGTT and HOMA-IR, reduce the serum levels of LDL-C, HDL-C, TG, and TC, restore neuronal dysfunction and synaptic plasticity, and decrease the deposition of Aβ in the hippocampus. The findings from ELISA, IF, and RT-qPCR revealed that JTW could alleviate microglial activation and inflammatory status in vivo and coptisine could play the same role in vitro. Moreover, the changes of the JAK2/STAT3 signaling pathway in LPS-induced BV2 cells or hippocampus of db/db mice were distinctly reversed by coptisine or JTW, respectively. Our study suggested that JTW and its effective component coptisine could alleviate diabetes mellitus-related cognitive impairment, closely linked to the JAK2/STAT3 signaling pathway. [Display omitted] [ABSTRACT FROM AUTHOR]

Jinmaitong (JMT) is a prescription of Traditional Chinese Medicine that is composed of 12 crude drugs. It has been used in the treatment of diabetic neuropathic pain (DNP) for more than 30 years. Microglia are thought to play an important role in neuropathic pain. This study aimed to evaluate the protective effect of JMT against DNP and to investigate the underlying mechanisms in which the microglia and JAK2/STAT3 signaling pathway were mainly involved. The chemical composition of JMT was analyzed using liquid chromatography tandem mass spectrometry. The diabetes model was constructed using 11 to 12-week-old male Zucker diabetic fatty (ZDF) rat (fa/fa). The model rats were divided into 5 groups and were given JMT at three dosages (11.6, 23.2, and 46.4 g/kg, respectively, calculated as the crude drug materials), JAK inhibitor AG490 (positive drug, 10 μg/day), and placebo (deionized water), respectively, for eight weeks (n = 6). Meanwhile, Zucker lean controls (fa/+) were given a placebo (n = 6). Body weight was tested weekly and blood glucose was monitored every 2 weeks. The mechanical allodynia and heat hyperalgesia were assessed using mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) tests. After treatment, the microglia activation marker Iba-1, CD11B, CD68, neuroinflammatory mediators, and mediators of the JAK2/STAT3 signaling pathway were compared between different groups. The mRNA and protein levels of target genes were assessed by quantitative real-time PCR and Western Blot, respectively. We found that JMT significantly inhibited the overactivation of microglia in spinal cords, and suppressed neuroinflammation of DNP model rats, thereby ameliorating neurological dysfunction and injuries. Furthermore, these effects of JMT could be attributed to the inhibition of the JAK2/STAT3 signaling pathway. Our findings suggested that JMT effectively ameliorated DNP by modulating microglia activation via inhibition of the JAK2/STAT3 signaling pathway. The present study provided a basis for further research on the therapeutic strategies of DNP. [Display omitted] • Traditional Chinese medicine Jinmaitong improves diabetic peripheral neuropathic pain in ZDF rats. • Jinmaitong inhibits microglia activation and regulates the neuroinflammatory factors and chemokines of ZDF rats. • Jinmaitong may exert its effects by inhibiting the JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

The aim of this study was to discover new biomarkers to detect breast cancer (BC), which is an aggressive cancer with a high mortality rate. In this study, bioinformatic analyses (differential analysis, weighted gene co-expression network analysis, and machine learning) were performed to identify potential candidate genes for BC to study their molecular mechanisms. Furthermore, Quantitative Real-time PCR and immunohistochemistry assays were used to examine the protein and mRNA expression levels of a particular candidate gene (DLGAP5). And the effects of DLGAP5 on cell proliferation, migration, invasion, and cell cycle were further assessed using the Cell Counting Kit-8 assay, colony formation, Transwell, wound healing, and flow cytometry assays. Moreover, the changes in the JAK2/STAT3 signaling-pathway-related proteins were detected by Western Blot. A total of 44 overlapping genes were obtained by differential analysis and weighted gene co-expression network analysis, of which 25 genes were found in the most tightly connected cluster. Finally, NEK2, CKS2, UHRF1, DLGAP5, and FAM83D were considered as potential biomarkers of BC. Moreover, DLGAP5 was highly expressed in BC. The down-regulation of DLGAP5 may inhibit the proliferation, migration, invasion, and cell cycle of BC cells, and the opposite was true for DLGAP5 overexpression. Correspondingly, silencing or overexpression of the DLGAP5 gene inhibited or activated the JAK2/STAT3 signaling pathway, respectively. DLGAP5, as a potential biomarker of BC, may impact the cell proliferation, migration, invasion, cell cycle, and BC development by modulating the JAK2/STAT3 signaling pathway.

T-cell lymphoma invasion and metastasis 2 (TIAM2) plays a critical role in the malignancy development of many human cancers. However, the specific regulatory mechanism of TIAM2 in osteosarcoma has not yet been explored. In this study, we investigated how TIAM2 affects the proliferation and invasion of osteosarcoma cells and the underlying molecular mechanism. We performed data mining of publicly available datasets to examine whether the expression of TIAM2 is associated with osteosarcoma. We knocked down the expression of and overexpressed TIAM2 in osteosarcoma cells. The proliferative capacity of cells in each group was determined by the Cell Counting Kit-8 assay. Wound healing and Transwell invasion assays were performed to evaluate the migration and invasion abilities of the TIAM2 knockdown and overexpressed osteosarcoma cells. Determination of the function of TIAM2 in vivo was performed in nude mice. Data mining confirmed that TIAM2 expression is associated with poor prognosis in osteosarcoma. TIAM2 expression levels were significantly higher in osteosarcoma cells, and TIAM2 expression knockdown reduced proliferation and invasion abilities. Animal experiments demonstrated that TIAM2 promotes tumor growth. Additional experiments suggested that TIAM2 was significantly related to the activation of the JAK2/STAT3 pathway, and subsequent mechanistic experiments further confirmed this. Our findings suggest that TIAM2 promotes the proliferation and invasion capacities of osteosarcoma cells by activating the JAK2/STAT3 signaling pathway. TIAM2 may serve as a potential prognostic target for patients with OS.