Levels Of Signal Transducer And Activator Of Transcription 3 Research Articles

Effects of an acetylcholinesterase inhibitor and an N-methyl-D-aspartate receptor antagonist on inflammation and degeneration of the nucleus pulposus.

The study aimed to examine the effects of two drugs, an acetylcholinesterase inhibitor (AChEI) and an N-methyl-D-aspartate receptor (NMDAR) antagonist, on degenerated annulus fibrosus (AF) and nucleus pulposus (NP) cells and the extracellular matrix (ECM) structure in vitro. Tissue samples were obtained from patients with intervertebral disc herniation (four males and four females; classified as Pfirmann stage IV) and used to prepare cell cultures. Untreated cell culture samples served as the control group. Study group samples were treated with donepezil, memantine or a combination of the two drugs. Cell viability, toxicity and proliferation were evaluated in all groups. Western blotting was used to examine changes in protein expression of signal transducer and activator of transcription 3 (STAT3), phospho-STAT3 (ser727), hypoxia-inducible factor (HIF)-1 alpha (HIF-1α) and nucleotide-binding oligomerisation domain (NOD) leucine-rich repeat (LRR)-containing proteins (NLR) family pyrin domain containing 3 (NLRP3) inflammasome. The alpha significance value was < 0.05. Analysis of the microscopy and commercial kit results revealed that cell proliferation was suppressed, and no cell death was observed. The protein expression levels of NLRP3, STAT3, ser727 and HIF-1α were lower in the samples treated with donepezil and memantine at 72 h (p < 0.05). The protein expression levels of NLRP3, STAT3, ser727 and HIF-1α were higher in the samples treated with the combination of donepezil and memantine (p < 0.05). The combined administration of memantine a NMDAR antagonist which can prevent neurodegeneration and donepezil an AChEI used for pain relief increased the protein expression levels in the anabolic pathway. However, it did not reduce the protein expression levels in the catabolic pathway. Therefore, further studies are needed to provide extensive insight into whether it may be among the potential targets for the therapy of intervertebral disc (IVD) diseases.

MiR-125b Inhibits Cell Proliferation and Induces Apoptosis in Human Colon Cancer SW480 Cells via Targeting STAT3.

Colon cancer is one of the most common types of cancer worldwide. Multiple studies have unveiled the key role of microRNAs (miRNAs) in the development of various types of cancer. However, the mechanism of action of miR-125b in the development and progression of colon cancer remains unknown. In this study, we explored the association of miR-125b and signal transducer and activator of transcription 3 (STAT3) and its role in the proliferation and apoptosis of SW480 colon cancer cells. The miR-125b expression in NCM460, SW480, HT29, and HCT8 cells was detected using quantitative real-time polymerase chain reaction (qRT-PCR). SW480 cells were transfected with lentiviruses of GFP-miR-125b and GFP-NC to establish a stable miR-125b overexpression colon cancer cell model and a control model. The targeting relationship between miR-125b and STAT3 was analyzed using bioinformatics and verified by the dual-luciferase reporter gene assay. Cell proliferation and apoptosis were assessed using the Cell Counting Kit-8 assay and TUNEL staining. The expression levels of STAT3, Bcl-2, and Bax were analyzed using Western blot analysis. It was found that the relative mRNA expression of miR-125b was decreased in SW480, HT29, and HCT8 cells compared with that in NCM460 cells (P<0.05). The luciferase reporter gene assay confirmed that miR-125b downregulated the STAT3 gene expression (P<0.05). Overexpression of miR-125b inhibited proliferation and promoted apoptosis in SW480 colon cancer cells and was accompanied by upregulated Bax expression and downregulated Bcl-2 expression (P<0.05). Re-expression of STAT3 promoted cell proliferation and inhibited cell apoptosis, whereas Bcl-2 expression increased, and Bax expression decreased (P<0.05). The miR-125b regulates the expression of Bax and Bcl-2 by downregulating the expression of STAT3, thereby inhibiting proliferation and inducing apoptosis of SW480 colon cancer cells.

Glaucocalyxin A alleviates lipopolysaccharide-induced inflammation and apoptosis in pulmonary microvascular endothelial cells and permeability injury by inhibiting STAT3 signaling.

Glaucocalyxin A (GLA), an ent-kauranoid diterpene derived from Rabdosia japonica var. glaucocalyx, possesses antibacterial, anti-oxidative and anti-neuroinflammatory properties. The present study aimed to investigate the potential mechanisms underlying GLA in the pathogenesis of pneumonia. Human pulmonary microvascular endothelial cells (hPMVECs) treated with lipopolysaccharide (LPS) were treated with GLA, followed by the detection of cell viability, inflammation, apoptosis and cell permeability. Furthermore, the protein expression levels of apoptosis- and permeability-associated proteins were determined using western blot analysis. Following treatment with a signal transducer and activator of transcription 3 (STAT3) activator, the protein expression levels of STAT3 and endoplasmic reticulum stress-associated proteins were determined, to confirm whether STAT3 signaling was mediated by GLA. Lastly, the mRNA expression level of inflammatory cytokines, apoptosis and permeability injury were also determined following treatment with a STAT3 activator. The results revealed that GLA ameliorated inflammation, apoptosis and permeability injury in LPS-induced hPMVECs. Following treatment with a STAT3 activator, the therapeutic effects of GLA on LPS-induced hPMVECs were abrogated. In conclusion, GLA alleviated LPS-induced inflammation, apoptosis and permeability injury in hPMVECs by inhibiting STAT3 signaling, which highlighted the potential therapeutic value of GLA in the treatment of pneumonia.

Association of STAT3, PTPRT, TNK2-AS1, LINC-ROR Genes Expression Level with Prostate Cancer and Benign Prostatic Hyperplasia

Background: Prostate cancer (PCa) and benign prostate hyperplasia (BPH) are highly prevalent heterogeneous disorders among men. Since angiogenesis is the key step in cancer progression, the deregulation of genes involved in this process may play a role in cancer development. Objectives: We evaluated the expression level of 4 angiogenesis-related genes including signal transducer and activator of transcription 3 (STAT3), protein tyrosine phosphatase receptor type T (PTPRT), TNK2 antisense RNA 1 (TNK2-AS1), and long intergenic non-protein coding rna-regulator of reprogramming (LINC-ROR) in patients with PCa and BPH. Methods: The expression level of STAT3, PTPRT, TNK2-AS1, and LINC-ROR genes in tumoral and adjacent non-cancerous tissue (ANCT) samples of 50 PCa patients and tissue samples from 50 BPH patients were evaluated, using the real-time PCR method. The statistical analysis was performed to evaluate the association between genes expression and clinicopathological characteristics of patients with PCa. Results: The expression level of STAT3 and LINC-ROR was upregulated in tumoral tissues compared to ANCTs (P &lt; 0.0001 for both). Only the expression level of STAT3 in PCa was higher than in BPH tissues (P = 0.001). The elevated expression of STAT3 was associated with the higher grade group of the tumor (P = 0.03). Also, the high expression level of PTPRT and LINC-ROR genes was associated with a higher stage of cancer in patients with PCa (P = 0.002, P = 0.0001 respectively). The STAT3 gene transcript level had an excellent diagnostic power for discrimination between tumoral tissue and the ANCTs with an area under the curve (AUC) of 0.93. Conclusions: The higher expression of STAT3 and LINC-ROR suggested a role in the pathogenesis of PCa in higher stages. Also, STAT3 expression level could be suggested as a potential biomarker for PCa in combination with PSA level.

Design, Synthesis and Structure-Activity Relationship Studies of Meridianin Derivatives as Novel JAK/STAT3 Signaling Inhibitors.

Hyperactivation of Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) signaling is an attractive therapeutic target for tumor therapy. Herein, forty-eight novel meridianin derivatives were designed and synthesized, and their antitumor activity was evaluated in vitro both for activity optimization and structure–activity relationship (SAR) study. The results indicated that most derivatives exhibited significantly improved antitumor activity, especially for compound 6e. The compound 6e contains an isothiouronium linked by an alkyl chain consisting of six carbon atoms with IC50 ranging from 1.11 to 2.80 μM on various cancer cell lines. Consistently, the 6e dose dependently induced the apoptosis of A549 and DU145 cells, in which STAT3 is constitutively active. Western blotting assays indicated that the phosphorylation levels of JAK1, JAK2 and STAT3 were inhibited by 6e at 5 μM without significant change in the total STAT3 level. Moreover, 6e also suppressed the expression of STAT3 downstream genes, including c-Myc, Cyclin D1 and Bcl-XL at 10 μM. An additional in vivo study revealed that 6e at the dose of 10 mg/kg could potently inhibit the DU145 xenograft tumor without obvious body weight loss. These results clearly indicate that 6e could be a potential antitumor agent by targeting the JAK/STAT3 signaling pathway.

Studying the Anticancer Properties of Bone Marrow-Derived Mesenchymal Stem Cells against Hepatocellular Carcinoma Induced by N-Nitrosodiethylamine in Male Rats

This study was initiated to emphasize the possibility that bone marrow-derived mesenchymal stem cells (BM-MSCs) could alleviate hepatocarcinogenesis in vivo. Forty rats were enrolled and distributed as follows; group 1 was set as a negative control, while all of groups 2, 3, and 4 were orally received N-nitrosodiethylamine (NDEA) for hepatocellular carcinoma induction (HCC). Group 2 was left untreated, whereas groups 3 and 4 were orally treated with BM-MSCs and doxorubicin (DOX), respectively. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and gamma-glutamyl transferase (γ-GT) activities were estimated. The study looked at serum alpha-fetoprotein (AFP), glypican-3 (GPC-3), signal transducer and activator of transcription 3 (STAT3), and suppressors of cytokine signaling 3 (SOCS3). The liver tissue sections were also examined histopathologically. Results: Serum AST, ALT, ALP, and γ-GT activity were all significantly higher in the HCC group, along with AFP, Gpc-3, and STAT3 levels associated with a significant reduction in SOCS3 level versus the negative control group. The current data indicated that BM-MSCs and DOX administration in HCC rats yielded a significant decline in serum levels of AFP, Gpc-3, and STAT3 along with significant enhancement in serum SOCS3 level. Histopathological examination revealed that treatment with BM-MSCs and DOX significantly reduced the disruption of hepatic tissue architecture caused by NDEA intoxication. Conclusion: The biochemical parameters and the structural organization of the liver were improved considerably in HCC groups treated with BM-MSCs or DOX. Finally, the current work demonstrated the anticancer activity of BM-MSCs against hepatocellular carcinoma in an experimental setting.

Total or Near-total Thyroidectomy in treatment of Thyroid Cancer.

Objectives:To observe the clinical efficacy on total or near-total thyroidectomy in the treatment of thyroid cancer.Methods:Ninety-four patients with thyroid cancer treated in Meiris Branch of the First Affiliated Hospital of Qiqihar Medical College from June 2018 to June 2020 were selected as subjects. According to different surgical methods, they were divided into observation group and control group, with 47 patients in each group. The control group was treated with total thyroidectomy, while the observation group received near-total thyroidectomy. The two groups were both followed up for one year. The therapeutic effect, surgery-related indexes (surgical duration, intraoperative bleeding volume, postoperative analgesia time and postoperative 24-h VAS score), the incidence of complications three months after surgery, and the serum levels of relevant indexes [parathyroid hormone (PTH), calcium ion (Ca2+) and signal transducer and activator of transcription 3 (STAT3)] before and 14 d after surgery were compared between the two groups. The difference in quality of life [the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30)] between the two groups one year after surgery was observed.Results:During the one year follow-up, there was no death in both groups. The total efficacy of the observation group was higher than that of the control group (0.05). The surgical duration, postoperative analgesia time and postoperative 24-h VAS score of the observation group were higher than those of the control group (p<0.05). However, no statistically significant difference was found in intraoperative bleeding volume between the two groups (p> 0.05). Three months after surgery, the total incidence of postoperative complications in the observation group was higher than that in the control group (p< 0.05). Fourteen days after surgery, the levels of PTH, Ca2+ and STAT3 in the two groups were lower than those before surgery, and the levels in the observation group were lower than those in the control group (p< 0.05). One year after surgery, cognitive, emotional, role, social and physical scores in the observation group were all lower than those in the control group, without statistically significant differences (p> 0.05).Conclusion:Total thyroidectomy is effective in the treatment of thyroid cancer, but has many postoperative complications. Clinicians need to choose the appropriate surgical method according to the actual condition of patients.

The Effect of STAT3 Signal Pathway Activation on Retinopathy of Prematurity.

Objective: To investigate the mechanism of activation of the signal transducer and activator of transcription 3 (STAT3) signal pathway in the process of retinopathy of prematurity (ROP).Methods: Sixty newborn Sprague-Dawley (SD) rats were randomly separated into the hyperoxia and air control groups (n = 30/in each group). The serum hepcidin level on 21 d was measured using the enzyme-linked immunosorbent assay (ELISA). The expression of HAMP and STAT3 protein in the liver was determined using reverse transcription-polymerase chain reaction (RT-PCR) and western blotting. Retinal neovasculature was evaluated by hematoxylin and eosin (HE) stain and fluorescein lectin. The retinal endothelial cells were treated with 250 μmol/L cobalt chloride for 72 h and added S3I-201. The STAT3 level was determined by western blotting.Results: The expression of STAT3 protein increased significantly after hyperoxia stimulation. The expression of HAMP mRNA in the hyperoxia group was significantly higher than that of the control group. The proliferation of retinal cells was inhibited, and the expression of STAT3 was increased. No significant difference was noted in vascular endothelial growth factor (VEGF) mRNA. The expression of STAT3 and VEGF mRNA was significantly reduced.Conclusion: The activation of the STAT3 signal pathway increased hepcidin expression, contributing to the pathogenesis of ROP. S3I-201 inhibited the expression of STAT3 and VEGF mRNA levels. This information provides potential novel therapeutic approach to the prevention and treatment of ROP.

IFIT3 (interferon induced protein with tetratricopeptide repeats 3) modulates STAT1 expression in small extracellular vesicles.

We have previously shown that the αvβ6 integrin plays a key role in promoting prostate cancer (PrCa) and it can be transferred to recipient cells via small extracellular vesicles (sEVs). Furthermore, we have reported in a proteomic analysis that αvβ6 integrin down-regulation increases the expression of IFIT3 (interferon induced protein with tetratricopeptide repeats 3) in PrCa cells and their derived sEVs. IFIT3 is a protein well known for being an antiviral effector, but recently its role in cancer has also been elucidated. To study the relationship between IFIT3 and STAT1 (signal transducer and activator of transcription 1), an upstream regulator of IFIT3, in PrCa cells and their released sEVs, we used CRISPR/Cas9 techniques to down-regulate the expression of the β6 integrin subunit, IFIT3 or STAT1. Our results show that IFIT3 and STAT1 are highly expressed in PrCa cells devoid of the β6 integrin subunit. However, IFIT3 but not STAT1, is present in sEVs derived from PrCa cells lacking the β6 integrin subunit. We demonstrate that loss of IFIT3 generates sEVs enriched in STAT1 but reduces the levels of STAT1 in the cells. As expected, IFIT3 is not detectable in STAT1 negative cells or sEVs. We thus propose that the observed STAT1 enrichment in sEVs is a compensatory mechanism for the loss of IFIT3. Overall, these results provide new insights into the intrinsic role of IFIT3 as a regulator of STAT1 expression in sEVs and in intercellular communication in PrCa.

Chromatin accessibility analysis identifies GSTM1 as a prognostic marker in human glioblastoma patients

BackgroundGlioblastoma (GBM) is a malignant human brain tumor that has an extremely poor prognosis. Classic mutations such as IDH (isocitrate dehydrogenase) mutations, EGFR (epidermal growth factor receptor) alternations, and MGMT (O6-methylguanine-methyltransferase) promoter hypermethylation have been used to stratify patients and provide prognostic significance. Epigenetic perturbations have been demonstrated in glioblastoma tumorigenesis. Despite the genetic markers used in the management of glioblastoma patients, new biomarkers that could predict patient survival independent of known biomarkers remain to be identified.MethodsATAC-seq (assay for transposase accessible chromatin followed by sequencing) and RNA-seq have been used to profile chromatin accessible regions using glioblastoma patient samples with short-survival versus long-survival. Cell viability, cell cycle, and Western blot analysis were used to characterize the cellular phenotypes and identify signaling pathways.ResultsAnalysis of chromatin accessibility by ATAC-seq coupled with RNA-seq methods identified the GSTM1 (glutathione S-transferase mu-1) gene, which featured higher chromatin accessibility in GBM tumors with short survival. GSTM1 was confirmed to be a significant prognostic marker to predict survival using a different GBM patient cohort. Knockdown of GSTM1 decreased cell viability, caused cell cycle arrest, and decreased the phosphorylation levels of the NF-kB (nuclear factor kappa B) p65 subunit and STAT3 (signal transducer and activator of transcription 3) (pSer727).ConclusionsThis report demonstrates the use of ATAC-seq coupled with RNA-seq to identify GSTM1 as a prognostic marker of GBM patient survival. Activation of phosphorylation levels of NF-kB p65 and STAT3 (pSer727) by GSTM1 is shown. Analysis of chromatin accessibility in patient samples could generate an independent biomarker that can be used to predict patient survival.

The Circadian Rhythms of STAT3 in the Rat Pineal Gland and Its Involvement in Arylalkylamine-N-Acetyltransferase Regulation.

In rodents, the melatonin production by the pineal gland is controlled through adrenergic signaling from the suprachiasmatic nuclei and regulation of the principal enzyme in its synthesis, arylalkylamine-N-acetyltransferase (AANAT). In the present study, we identified increased isoprenaline-induced aa-nat expression and nocturnal AANAT activity in the pineal glands in response to the silencing of the signal transducer and activator of transcription 3 (STAT3) with siRNA or STAT3 inhibitors WP1066 and AZD1480. This AANAT activity enhancement in vivo did not interfere with light-induced AANAT suppression. Systemic or in vitro lipopolysaccharide (LPS) administration markedly increased Stat3 expression and STAT3 phosphorylation, but it did not significantly affect AANAT expression or activity. Simultaneous LPS administration and Stat3 silencing enhanced the aa-nat transcription and AANAT activity to a similar extent as Stat3 inhibition without LPS co-administration. Furthermore, we describe the circadian rhythmicity in Stat3 expression and the phosphorylated form of STAT3 protein in the rat pineal gland. Our data suggest that the higher nocturnal endogenous level of STAT3 in the pineal gland decelerates or hampers the process of NA-induced AANAT activation or affects the AANAT enzyme stability.

DNA methylation in promoter region of immune related genes STAT3 and VEGFA and biochemical parameters change in muscle of Japanese flounder under acute hypoxia

Acute hypoxic stress can lead to immune response in fish, but the molecular mechanism of muscle immunity in fish under acute hypoxia are still unclear. In this study, we carried out the effect of signal transducer and activator of transcription3(STAT3) and vascular endothelial growth factor A(VEGFA) on muscle immune responses of Japanese flounder (Paralichthys olivaceus) during acute hypoxic stimulation (1.65 ± 0.28mg/L O2; 3h, 6h, 12h, 24h) and reoxygenation (7.30 ± 0.40mg/L O2; R12h, R24h, R48h). In situ hybridization (ISH) showed that STAT3 and VEGFA RNA were co-located in the skeletal muscle of Japanese flounder. Japanese flounder was seriously affected by hypoxia for 3h and 6h. The expression of STAT3 and VEGFA increased significantly. The methylation levels of STAT3 5′UTR region and VEGFA promoter region were significantly lower than those in normoxia group, which was negatively correlated with the expression levels of STAT3 and VEGFA. The enzyme activities (LDH, ALT, AST, ALP) changed significantly. In addition, enzyme-linked immunosorbent assay (ELISA) detected a positive correlation between serum VEGFA concentration and muscle VEGFA mRNA. The current study have shown that Japanese flounder responded to acute hypoxic stress at multiple metabolic levels by changing DNA methylation status and activating transcription factors such as HIF-1α, Nrf2 and STAT3. It is significant for the scientific development of aquaculture through analyzing the effects of hypoxia on biological immunity.

SDF-1/CXCR4 axis promotes osteogenic differentiation of BMSCs through the JAK2/STAT3 pathway.

This study aimed to investigate the effects of stromal cell-derived factor-1 (SDF-1) and activation of its receptor, chemokine receptor 4 (CXCR4), on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and the key signaling mechanisms involved in these effects. BMSCs were treated with 100 μg/L SDF-1 and cultured in osteogenic medium for 7 days. RT-qPCR and western blotting were used to detect the protein and mRNA levels of Janus kinase 2 (JAK2), signal transducer and activator of transcription 3 (STAT3), Runt-related transcription factor 2 (Runx2), and osteocalcin (OCN). Alizarin-red staining was used to detect the mineralization-inducing ability of the cells. After BMSCs were treated with SDF-1, the levels of JAK2 mRNA, STAT3 mRNA, and protein phosphorylation increased, the number of mineralized nodules of BMSCs increased, and the osteogenic-differentiation ability was enhanced. In addition, after BMSCs were treated with an inhibitor of JAK2 phosphorylation, the levels of JAK2, STAT3, Runx2, and OCN decreased significantly, the number of mineralized nodules of BMSCs also decreased, and the osteogenic-differentiation ability decreased. The inhibition of CXCR4-treated BMSCs further confirmed that SDF-1/CXCR4 activated JAK2/STAT3 to regulate the osteogenic differentiation of BMSCs. SDF-1/CXCR4 promoted the osteogenic differentiation of BMSCs through JAK2/STAT3 activation.

Dihydrotanshinone I inhibits the growth of hepatoma cells by direct inhibition of Src

BackgroundLiver cancer is one of the leading causes of cancer-related death worldwide. Dihydrotanshinone I (DHI) was shown to inhibit the growth of several types of cancer. However, research related to hepatoma treatment using DHI is limited. PurposeHere, we explored the inhibitory effect of DHI on the growth of hepatoma cells, and investigated the underlying molecular mechanisms. MethodsThe proliferation of Hep3B, SMCC-7721 and SK-Hep1 hepatoma cells was evaluated using the MTS and Edu staining assay. Hepatoma cell death was analyzed with a LIVE/DEAD Cell Imaging Kit. The relative expression and phosphorylation of proto-oncogene tyrosine-protein kinase Src (Src) and signal transducer and activator of transcription-3 (STAT3) proteins in hepatoma cells, as well as the expression of other protein components, were measured by western blotting. The structural interaction of DHI with Src proteins was evaluated by molecular docking, molecular dynamics simulation, surface plasmon resonance imaging and Src kinase inhibition assay. Src overexpression was achieved by infection with an adenovirus vector encoding human Src. Subsequently, the effects of DHI on tumor growth inhibition were further validated using mouse xenograft models of hepatoma. ResultsIn vitro studies showed that treatment with DHI inhibited the proliferation and promoted cell death of Hep3B, SMCC-7721 and SK-Hep1 hepatoma cells. We further identified and verified Src as a direct target of DHI by using molecular stimulation, surface plasmon resonance image and Src kinase inhibition assay. Treatment with DHI reduced the in vitro phosphorylation levels of Src and STAT3, a transcription factor regulated by Src. In the xenograft mouse models, DHI dose-dependently suppressed tumor growth and Src and STAT3 phosphorylation. Moreover, Src overexpression partly abrogated the inhibitory effects of DHI on the proliferation and cell death in hepatoma cells. ConclusionOur results suggest that DHI inhibits the growth of hepatoma cells by direct inhibition of Src.

Interference with the expression of S1PR1 or STAT3 attenuates valvular damage due to rheumatic heart disease.

Rheumatic heart disease (RHD) affects numerous individuals annually; however, its pathogenesis remains unclear. The sphingosine 1-phosphate receptor 1 (S1PR1) and signal transducer and activator of transcription 3 (STAT3) have recently been shown to be involved in valvular damage via the promotion of the differentiation of T helper 17 (Th17) cells during the development of RHD-induced valvular damage. The present study investigated whether altering the expression of S1PR1 or STAT3 attenuates valvular damage due to RHD. Inactivated group A streptococcus (GAS) was used to establish a rat model of RHD. Recombinant adeno-associated viral vectors carrying an S1PR1 overexpression sequence were used to overexpress S1PR1. STAT3 small interfering RNA (STAT3-siRNA) was used to inhibit STAT3 expression. Reverse transcription-quantitative PCR (RT-qPCR) was performed to detect the mRNA expression of S1PR1, STAT3, collagen type III α1 chain (Col3a1) and fibroblast-specific protein 1. Western blotting (WB) and immunohistochemistry were used to detect the levels of S1PR1, STAT3, phosphorylated (p-) STAT3, and retinoic acid-related orphan receptor γT (RORγt) proteins. Enzyme-linked immunosorbent assays (ELISAs) and immunohistochemistry were used to detect the levels of interleukin (IL)-6 and IL-17. Hematoxylin and eosin (H&E) staining and Sirius Red staining were performed to evaluate the degree of inflammation and fibrosis in the valvular tissues. S1PR1 expression was decreased in the valvular tissues of the rats with RHD. The levels of IL-6, IL-17 and p-STAT3 in the rats with RHD were increased. The degree of valvular inflammation and fibrosis in the rats with RHD was also increased. The overexpression of S1PR1 and the inhibition of STAT3 reduced the total p-STAT3 level, resulting in decreased levels of IL-6, IL-17 and RORγt, and a reduced degree of valvular inflammation and fibrosis. These results suggest that the expression of S1PR1 and STAT3 may be involved in valvular tissue damage due to RHD. Thus, strategies designed to interfere with the expression of S1PR1 or STAT3 may affect the expression of Th17 cell-related cytokines and may thus attenuate valvular damage due to RHD.

Diospyros Peregrina Fruit Preparation Mediated Immunomodulation of Lymphocytes Isolated From The Blood of Breast Cancer Patients.

Breast cancer is an uncontrolled growth of epithelial cells. The loss of Breast Cancer gene1 (BRCA1) activity due to mutation or down-regulation of gene expression promotes tumorigenesis and increases the risk of breast cancer. We aimed to pulsate lymphocytes of breast cancer patients and normal individuals, using Diospyros peregrina fruit preparation (DFP) to study the cancer-protective immunity, and the signal transduction processes involved with it. We also investigated the role of DFP in the release of lymphocytic nitric oxide (NO), which is a key tumoricidal agent, known to regulate T-cell proliferation, cytokine production, cell signaling, and apoptosis. Using Ficoll-Hypaque gradient centrifugation, lymphocytes were isolated from the blood of 12 patients and 12 normal individuals. Cells were treated with or without DFP (2.5 µg/ml) for 48 hours. Both non-stimulated and stimulated cells were then subjected to MTT and NO release assay; following which qPCR was performed to estimate mRNA levels and percentage enrichment of certain genes. DFP stimulates lymphocytic proliferation (p=0.0118) and release of NO (p=0.01) significantly. DFP also noticeably enhances the expression of T helper (TH) cell 1 specific interferon-gamma (IFNG), interleukin 12 (IL12), T-Box Transcription Factor 21 (TBX21) and signal transducer and activator of transcription 1 (STAT1) genes. DFP treatment significantly increases tumor protective immunity by decreasing the expression levels of TH2 network-specific GATA3 and interleukin 4 (IL4) genes but increasing the expression levels of TH1 network-specific IFNG, IL12, TBX21, and STAT1 genes. DFP increases the expression levels of TH1 specific network genes which in turn help in evoking tumor protective immunity.

Effects of manganese on microglia M1/M2 polarization and SIRT1‐mediated transcription of STAT3‐dependent genes in mouse

Overexposure to manganese (Mn) can lead to neurological diseases, characterized by behavioral and motor impairments. Microglia-mediated neuroinflammation is involved in the pathogenesis and progression of neurodegenerative diseases. Microglial M1 and M2 phenotypes play pro-inflammatory and anti-inflammatory roles, respectively, in response to microenvironmental disturbances. Silent information regulator (SIRT1) has been demonstrated to play an important role in the neuroinflammatory response by deacetylating various transcription factors, such as proliferator-activated receptor γ coactivator 1α (PGC-1α), nuclear factor kappa B (NF-κB), and signal transducer and activator of transcription 3 (STAT3). In addition, PGC-1α and STAT3 have been implicated in microglial polarization and inflammatory response. In this study, Mn exposure (50, 100, 200μmol/kg) induced neuroinflammatory injury and interfered with microglial M1/M2 polarization in mice, as indicated by the upregulated expression of M1 polarization marker mRNA (IL-1β, IL-6, TNF-α, and iNOS), whereas changes in M2 polarization markers (IL-4, TGF-β, and Arg1) varied, with some increasing and some decreasing, in response to increasing Mn doses, which was consistent with the flow cytometry results used to detect the percentages of each microglial type. We found that Mn could downregulate SIRT1 expression and activate NF-κB signaling. Following mice in the 200 μmol/kg Mn treatment, STAT3 and PGC-1α levels in the nuclear fraction both significantly decreased, and the interaction between the proteins decreased, affecting the transcription of STAT3-mediated genes. These findings provide new insights regarding the role played by Mn neurotoxicity in the suppression of neuroinflammation through the regulation of microglial polarization.

Periplogenin suppresses the growth of esophageal squamous cell carcinoma in vitro and in vivo by targeting STAT3.

The mortality rate of esophageal squamous cell carcinoma (ESCC) is higher than that of other cancers worldwide owing to a lack of therapeutic targets and related drugs. This study aimed to find new drugs by targeting an efficacious therapeutic target in ESCC patients. Signal transducer and activator of transcription 3 (STAT3) is hyperactive in ESCC. Herein, we identified a novel STAT3 inhibitor, periplogenin, which strongly inhibited phosphorylation of STAT3 at Tyr705. Docking models and pull-down assays revealed that periplogenin bound directly and specifically to STAT3, leading to significant suppression of subsequent dimerization, nuclear import, and transcription activities. In addition, STAT3 knockdown cell lines were insensitive to periplogenin, whereas in contrast, STAT3-overexpressing cells were more sensitive to periplogenin, indicating that STAT3 was a target of periplogenin. Intraperitoneally administered periplogenin exhibited efficacious therapeutic effects in ESCC patient-derived xenograft models and dramatically impaired the phosphorylation of STAT3 and expression levels of STAT3-mediated downstream genes. Thus, our study demonstrated that periplogenin acted as a new STAT3 inhibitor, suppressing the growth of ESCC in vitro and in vivo, providing a basis for its potential application in ESCC treatment and prevention.

Hsa_circ_0001610 knockdown modulates miR-646-STAT3 axis to suppress endometrial carcinoma progression.

Endometrial carcinoma (EC) development is associated with dysregulated circular RNA profiles. The purpose of the current research is to study the role and mechanism of hsa_circ_0001610 (circ_0001610) in EC progression. circ_0001610, microRNA (miR)-646, and signal transducer and activator of transcription 3 (STAT3) expression levels were measured in EC. Functional analyses were performed using Cell Counting Kit-8, colony formation, transwell, wound healing, flow cytometry, glycolysis, and xenograft analyses. Binding association was evaluated with dual-luciferase reporter assay. circ_0001610 levels were upregulated in EC samples (n = 30) and cells. circ_0001610 interference restrained cell proliferation, migration, and invasion, and promoted apoptosis. circ_0001610 downregulation constrained glycolysis through reducing glucose consumption, lactate production, and levels of adenosine triphosphate, extracellular acidification, hexokinase 2, and lactate dehydrogenase A, and increasing oxygen consumption rate. miR-646 is targeted by circ_0001610, and miR-646 inhibition attenuated interference of circ_0001610-mediated suppression of EC development. STAT3 was modulated by miR-646, and miR-646 upregulation restrained EC progression by decreasing STAT3. circ_0001610 silencing reduced STAT3 levels by sponging miR-646 and reduced the growth of xenograft tumor established by EC cells. circ_0001610 knockdown represses EC progression through modulating the miR-646-STAT3 axis.

Upregulation of KDM6B contributes to lipopolysaccharide-induced anxiety-like behavior via modulation of VGLL4 in mice

Histone H3K27me3 demethylase KDM6B (also known as Jumonji domain-containing protein D3, JMJD3) plays vital roles in the etiology of inflammatory responses; however, little is known about the role of KDM6B in neuroinflammation-induced anxiety-like behavior. The present study aimed to investigate the potential role of KDM6B in lipopolysaccharide (LPS)-induced anxiety-like behavior and to evaluate whether it is associated with the modulation of vestigial-like family member 4 (VGLL4). The elevated plus maze, light-dark box, and open-field test were performed to test the anxiety-like behavior induced by LPS in C57BL/6 J male mice. Levels of relative protein expression in the hippocampus were quantified by western blotting. KDM6B inhibitor GSK-J4 and microglia inhibitor minocycline as well as adeno-associated virus of Vgll4 shRNA were used to explore the underlying mechanisms. We found that KDM6B, VGLL4, interleukin-1β (IL-1β), and ionized calcium-binding adaptor molecule-1 (Iba-1, microglia marker) protein levels were increased in LPS-dose dependent manner in the hippocampus but not in prefrontal cortex. GSK-J4 treatment attenuated LPS-induced VGLL4, the signal transducer and activator of transcription 3 (STAT3), IL-1β and Iba-1 upregulation and anxiety-like behavior. Knockdown VGLL4 with Vgll4 shRNA prevented the increase of anxiety-like behavior and levels of STAT3, IL-1β, and Iba-1 expression in the hippocampus of LPS-treated mice. Moreover, minocycline, an inhibitor of microglia treatment blunted LPS-induced anxiety-like behavior. Collectively, these results demonstrate that the induction of neuroinflammation by LPS promotes KDM6B activation in the hippocampus, and LPS-induced anxiety-like behavior is associated with upregulation of VGLL4 by KDM6B in the hippocampus.