Effects of interleukin-10 treated macrophages on bone marrow mesenchymal stem cells via signal transducer and activator of transcription 3 pathway

BACKGROUND Alveolar bone defects caused by inflammation are an urgent issue in oral implant surgery that must be solved.Regulating the various phenotypes of macrophages to enhance the inflammatory environment can significantly affect the progression of diseases and tissue engineering repair process.AIM To assess the influence of interleukin-10(IL-10)on the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)following their interaction with macrophages in an inflammatory environment.METHODS IL-10 modulates the differentiation of peritoneal macrophages in Wistar rats in an inflammatory environment.In this study,we investigated its impact on the proliferation,migration,and osteogenesis of BMSCs.The expression levels of signal transducer and activator of transcription 3(STAT3)and its activated form,phos-phorylated-STAT3,were examined in IL-10-stimulated macrophages.Subsequently,a specific STAT3 signaling inhibitor was used to impede STAT3 signal activation to further investigate the role of STAT3 signaling.RESULTS IL-10-stimulated macrophages underwent polarization to the M2 type through substitution,and these M2 macrophages actively facilitated the osteogenic differentiation of BMSCs.Mechanistically,STAT3 signaling plays a crucial role in the process by which IL-10 influences macrophages.Specifically,IL-10 stimulated the activation of the STAT3 signaling pathway and reduced the macrophage inflammatory response,as evidenced by its diminished impact on the osteogenic differentiation of BMSCs.CONCLUSION Stimulating macrophages with IL-10 proved effective in improving the inflammatory environment and promoting the osteogenic differentiation of BMSCs.The IL-10/STAT3 signaling pathway has emerged as a key regulator in the macrophage-mediated control of BMSCs’osteogenic differentiation.

Diffusion-weighted magnetic resonance imaging reflects activation of signal transducer and activator of transcription 3 during focal cerebral ischemia/reperfusion

Signal transducer and activator of transcription（STAT）is a unique protein family that binds to DNA,coupled with tyrosine phosphorylation signaling pathways,acting as a transcriptional regulator to mediate a variety of biological effects.Cerebral ischemia and reperfusion can activate STATs signaling pathway,but no studies have confirmed whether STAT activation can be verified by diffusion-weighted magnetic resonance imaging（DWI）in rats after cerebral ischemia/reperfusion.Here,we established a rat model of focal cerebral ischemia injury using the modified Longa method.DWI revealed hyperintensity in parts of the left hemisphere before reperfusion and a low apparent diffusion coefficient.STAT3 protein expression showed no significant change after reperfusion,but phosphorylated STAT3 expression began to increase after 30 minutes of reperfusion and peaked at 24 hours.Pearson correlation analysis showed that STAT3 activation was correlated positively with the relative apparent diffusion coefficient and negatively with the DWI abnormal signal area.These results indicate that DWI is a reliable representation of the infarct area and reflects STAT phosphorylation in rat brain following focal cerebral ischemia/reperfusion.

18β-glycyrrhetinic acid promotes gastric cancer cell autophagy and inhibits proliferation by regulating miR-328-3p/signal transducer and activator of transcription 3

BACKGROUND Gastric cancer(GC)is one of the most common cancer types worldwide,and its prevention and treatment methods have garnered much attention.As the active ingredient of licorice,18β-glycyrrhetinic acid(18β-GRA)has a variety of pharmacological effects.The aim of this study was to explore the effective target of 18β-GRA in the treatment of GC,in order to provide effective ideas for the clinical prevention and treatment of GC.AIM To investigate the mechanism of 18β-GRA in inhibiting cell proliferation and promoting autophagy flux in GC cells.METHODS Whole transcriptomic analyses were used to analyze and screen differentially expressed microRNAs(miRNAs)in GC cells after 18β-GRA intervention.Lentivirus-transfected GC cells and the Cell Counting Kit-8 were used to detect cell proliferation ability,cell colony formation ability was detected by the clone formation assay,and flow cytometry was used to detect the cell cycle and apoptosis.A nude mouse transplantation tumor model of GC cells was constructed to verify the effect of miR-328-3p overexpression on the tumorigenicity of GC cells.Tumor tissue morphology was observed by hematoxylin and eosin staining,and microtubule-associated protein light chain 3(LC3)expression was detected by immunohistochemistry.TransmiR,STRING,and miRWalk databases were used to predict the relationship between miR-328-3p and signal transducer and activator of transcription 3(STAT3)-related information.Expression of STAT3 mRNA and miR-328-3p was detected by quantitative polymerase chain reaction(qPCR)and the expression levels of STAT3,phosphorylated STAT3(p-STAT3),and LC3 were detected by western blot analysis.The targeted relationship between miR-328-3p and STAT3 was detected using the dual-luciferase reporter gene system.AGS cells were infected with monomeric red fluorescent protein-green fluorescent protein-LC3 adenovirus double label.LC3 was labeled and autophagy flow was observed under a confocal laser microscope.RESULTS The expression of miR-328-3p was significantly upregulated after 18β-GRA intervention in AGS cells(P=4.51E-06).Overexpression of miR-328-3p inhibited GC cell proliferation and colony formation ability,arrested the cell cycle in the G0/G1 phase,promoted cell apoptosis,and inhibited the growth of subcutaneous tumors in BALB/c nude mice(P<0.01).No obvious necrosis was observed in the tumor tissue in the negative control group(no drug intervention or lentivirus transfection)and vector group(the blank vector for lentivirus transfection),and more cells were loose and necrotic in the miR-328-3p group.Bioinformatics tools predicted that miR-328-3p has a targeting relationship with STAT3,and STAT3 was closely related to autophagy markers such as p62.After overexpressing miR-328-3p,the expression level of STAT3 mRNA was significantly decreased(P<0.01)and p-STAT3 was downregulated(P<0.05).The dual-luciferase reporter gene assay showed that the luciferase activity of miR-328-3p and STAT33’untranslated regions of the wild-type reporter vector group was significantly decreased(P<0.001).Overexpressed miR-328-3p combined with bafilomycin A1(Baf A1)was used to detect the expression of LC3 II.Compared with the vector group,the expression level of LC3 II in the overexpressed miR-328-3p group was downregulated(P<0.05),and compared with the Baf A1 group,the expression level of LC3 II in the overexpressed miR-328-3p+Baf A1 group was upregulated(P<0.01).The expression of LC3 II was detected after intervention of 18β-GRA in GC cells,and the results were consistent with the results of miR-328-3p overexpression(P<0.05).Additional studies showed that 18β-GRA promoted autophagy flow by promoting autophagosome synthesis(P<0.001).qPCR showed that the expression of STAT3 mRNA was downregulated after drug intervention(P<0.05).Western blot analysis showed that the expression levels of STAT3 and p-STAT3 were significantly downregulated after drug intervention(P<0.05).CONCLUSION 18β-GRA promotes the synthesis of autophagosomes and inhibits GC cell proliferation by regulating the miR-328-3p/STAT3 signaling pathway.

Regulation and function of signal transducer and activator of transcription 3

Signal transducer and activator of transcription 3(STAT3), a member of the STAT family, is a key regulator of many physiological and pathological processes. Significant progress has been made in understanding the transcriptional control, posttranslational modification, cellular localization and functional regulation of STAT3. STAT3 can translocate into the nucleus and bind to specific promoter sequences, thereby exerting transcriptional regulation. Recent studies have shown that STAT3 can also translocate into mitochondria, participating in aerobic respiration and apoptosis. In addition, STAT3 plays an important role in inflammation and tumorigenesis by regulating cell proliferation, differentiation and metabolism. Conditional knockout mouse models make it possible to study the physiological function of STAT3 in specific tissues and organs. This review summarizes the latest advances in the understanding of the expression, regulation and function of STAT3 in physiological and tumorigenic processes.

Signal transducer and activator of transcription 3 promotes the Warburg effect possibly by inducing pyruvate kinase M2 phosphorylation in liver precancerous lesions

BACKGROUND Study shows that signal transducer and activator of transcription 3(STAT3) can increase the Warburg effect by stimulating hexokinase 2 in breast cancer and upregulate lactate dehydrogenase A and pyruvate dehydrogenase kinase 1 in myeloma. STAT3 and pyruvate kinase M2(PKM2) can also be activated and enhance the Warburg effect in hepatocellular carcinoma. Precancerous lesions are critical to human and rodent hepatocarcinogenesis. However, the underlying molecular mechanism for the development of liver precancerous lesions remains unknown. We hypothesized that STAT3 promotes the Warburg effect possibly by upregulating p-PKM2 in liver precancerous lesions in rats.AIM To investigate the mechanism of the Warburg effect in liver precancerous lesions in rats.METHODS A model of liver precancerous lesions was established by a modified Solt-Farber method. The liver pathological changes were observed by HE staining and immunohistochemistry. The transformation of WB-F344 cells induced with Nmethyl-N'-nitro-N-nitrosoguanidine and hydrogen peroxide was evaluated by the soft agar assay and aneuploidy. The levels of glucose and lactate in the tissue and culture medium were detected with a spectrophotometer. The protein levels of glutathione S-transferase-π, proliferating cell nuclear antigen(PCNA), STAT3,and PKM2 were examined by Western blot and immunofluorescence.RESULTS We found that the Warburg effect was increased in liver precancerous lesions in rats. PKM2 and p-STAT3 were upregulated in activated oval cells in liverprecancerous lesions in rats. The Warburg effect, p-PKM2, and p-STAT3 expression were also increased in transformed WB-F344 cells. STAT3 activation promoted the clonal formation rate, aneuploidy, alpha-fetoprotein expression,PCNA expression, G1/S phase transition, the Warburg effect, PKM2 phosphorylation, and nuclear translocation in transformed WB-F344 cells.Moreover, the Warburg effect was inhibited by stattic, a specific inhibitor of STAT3, and further reduced in transformed WB-F344 cells after the intervention for PKM2.CONCLUSION The Warburg effect is initiated in liver precancerous lesions in rats. STAT3 activation promotes the Warburg effect by enhancing the phosphorylation of PKM2 in transformed WB-F344 cells.

Downregulation of signal transducer and activator of transcription 3 by sorafenib:A novel mechanism for hepatocellular carcinoma therapy

Hepatocellular carcinoma is one of the most common cancers worldwide,and a leading cause of cancer-related death.Owing to unsatisfactory clinical outcomes under the current standard of care,there is a need to search for and identify novel and potent therapeutic targets to improve patient outcomes.Sorafenib is the first and only approved targeted therapy for the treatment of hepatocellular carcinoma.Besides functioning as a multiple tyrosine kinase,sorafenib also acts via a kinase-independent mechanism to target signal transducer and activator of transcription 3(STAT3) signaling in hepatocellular carcinoma cells.STAT3 is a key regulator of inflammation,cell survival,and tumorigenesis of liver cells,and the high percentage of hepatocellular carcinoma cells with constitutively active STAT3 justifies targeting it for the development of novel therapeutics.Sorafenib inactivates STAT3 and STAT3-related signaling by inducing a conformational change in and releasing the autoinhibition of Src homology region 2 domaincontaining phosphatase-1.This phosphatase negatively regulates STAT3 activity,which leads to the subsequent apoptosis of cancer cells.The novel anti-cancer property of sorafenib will be discussed in this review,not only adding information regarding its mechanism of action but also providing an innovative approach for the development of cancer therapeutics in the future.

Hepatitis C Virus non-structural 5A abrogates signal transducer and activator of transcription-1 nuclear translocation induced by IFN-α through dephosphorylation

AIM： To study the effect of Hepatitis C virus nonstructural 5A （HCV NSSA） on IFNα induced signal transducer and activator of transcription-1 （STAT1） phosphorylation and nuclear translocation.METHODS： Expression of STAT1 Tyr701 phosphorylation at different time points was confirmed by Western blot, and the time point when p-STAT1 expressed most, was taken as the IFN induction time for further studies. Immunocytochemistry was used to confirm the successful transient transfection of NS5A expression plasmid. Immunofluorescene was performed to observe if there was any difference in IFNα-induced STAT1 phosphorylation and nuclear translocation between HCV NSSA-expressed and non-HCV NSSA-expressed cells. Western blot was used to compare the phosphorylated STAT1 protein of the cells.RESULTS： Expression of HCV NS5A was found in the cytoplasm of pCNS5A-transfected Huh7 cells, but not in the PRC/ CMV transfected or non-transfected cells, STAT1 Tyr701 phosphorylation was found strongest in 30 min of IFN induction, STAT1 phosphorylation and nuclear import were much less in the presence of HCV NS5A protein in contrast to pRC/CMV-transfected and non-transfected cells under fluorescent microscopy, which was further confirmed by Western blot.CONCLUSION： HCV NSSA expression plasmid is successfully transfected into Huh7 cells and HCV NS5A protein is expressed in the cytoplasm of the cells. IFN-α is able to induce STAT1 phosphrylation and nuclear translocation, and this effect is inhibited by HCV NS5A protein, which might be another possible resistance mechanism to interferon alpha therapy.

Apigenin ameliorates imiquimod-induced psoriasis in C57BL/6J mice by inactivating STAT3 and NF-κB

Psoriasis is a chronic autoimmune disease featured by patches on the skin.It is caused by malfunction of immune cells and keratinocytes with inflammation as one of its key features.Apigenin(API)is a natural flavonoid with anti-inflammatory and immunoregulatory properties.Therefore,we speculated that API can ameliorate psoriasis,and determined its effect on the development of psoriasis by using imiquimod(IMQ)-induced psoriasis mouse model.Our results showed that API attenuated IMQ-induced phenotypic changes,such as erythema,scaling and epidermal thickening,and improved splenic hyperplasia.Abnormal differentiation of immune cells was restored in API-treated mice.Mechanistically,we revealed that API is a key regulator of signal transducer activator of transcription 3(STAT3).API regulated immune responses by reducing interleukin-23(IL-23)/STAT3/IL-17A axis.Moreover,it suppressed IMQ-caused cell hyperproliferation by inactivating STAT3 through regulation of extracellular signal-regulated kinase 1/2 and nuclear factor-κB(NF-κB)pathway.Furthermore,API reduced expression of inflammatory cytokines through inactivation of NF-κB.Taken together,our study demonstrates that API can ameliorate psoriasis and may be considered as a strategy for psoriasis treatment.

STAT3-Dependent Effects of Polymeric Immunoglobulin Receptor in Regulating Interleukin-17 Signaling and Preventing Autoimmune Hepatitis

One-third of patients with autoimmune hepatitis(AIH)have cirrhosis at the time of diagnosis.The relevance of these variables,although unknown,is believed to be critical in AIH because of suspected interactions between the gut microbiome and genetic factors.Dysbiosis of the gut flora and elevated polymeric immunoglobulin receptor(pIgR)levels have been observed in both patients and mouse models.Moreover,there is a direct relationship between pIgR expression and transaminase levels in patients with AIH.In this study,we aimed to explore how pIgR influences the secretion of regenerating islet-derived 3 beta(Reg3b)and the flora composition in AIH using in vivo experiments involving patients with AIH and a concanavalin A-induced mouse model of AIH.Reg3b expression was reduced in pIgR gene(Pigr)-knockout mice compared to that in wild-type mice,leading to increased microbiota disruption.Conversely,exogenous pIgR supplementation increased Reg3b expression and maintained microbiota homeostasis.RNA sequencing revealed the participation of the interleukin(IL)-17 signaling pathway in the regulation of Reg3b through pIgR.Furthermore,the introduction of external pIgR could not restore the imbalance in gut microbiota in AIH,and the decrease in Reg3b expression was not apparent following the inhibition of signal transducer and activator of transcription 3(STAT3).In this study,pIgR facilitated the upregulation of Reg3b via the STAT3 pathway,which plays a crucial role in preserving the balance of the intestinal microbiota in AIH.Through this research,we discovered new molecular targets that can be used for the diagnosis and treatment of AIH.

Gossypol acetic acid regulates leukemia stem cells by degrading LRPPRC via inhibiting IL-6/JAK1/STAT3 signaling or resulting mitochondrial dysfunction

BACKGROUND Leukemia stem cells(LSCs)are found to be one of the main factors contributing to poor therapeutic effects in acute myeloid leukemia(AML),as they are protected by the bone marrow microenvironment(BMM)against conventional therapies.Gossypol acetic acid(GAA),which is extracted from the seeds of cotton plants,exerts anti-tumor roles in several types of cancer and has been reported to induce apoptosis of LSCs by inhibiting Bcl2.AIM To investigate the exact roles of GAA in regulating LSCs under different microenvironments and the exact mechanism.METHODS In this study,LSCs were magnetically sorted from AML cell lines and the CD34+CD38-population was obtained.The expression of leucine-rich pentatricopeptide repeat-containing protein(LRPPRC)and forkhead box M1(FOXM1)was evaluated in LSCs,and the effects of GAA on malignancies and mitochondrial RESULTS LRPPRC was found to be upregulated,and GAA inhibited cell proliferation by degrading LRPPRC.GAA induced LRPPRC degradation and inhibited the activation of interleukin 6(IL-6)/janus kinase(JAK)1/signal transducer and activator of transcription(STAT)3 signaling,enhancing chemosensitivity in LSCs against conventional chemotherapies,including L-Asparaginase,Dexamethasone,and cytarabine.GAA was also found to downregulate FOXM1 indirectly by regulating LRPPRC.Furthermore,GAA induced reactive oxygen species accumulation,disturbed mitochondrial homeostasis,and caused mitochondrial dysfunction.By inhibiting IL-6/JAK1/STAT3 signaling via degrading LRPPRC,GAA resulted in the elimination of LSCs.Meanwhile,GAA induced oxidative stress and subsequent cell damage by causing mitochondrial damage.CONCLUSION Taken together,the results indicate that GAA might overcome the BMM protective effect and be considered as a novel and effective combination therapy for AML.

Galectin 2 regulates JAK/STAT3 signaling activity to modulate oral squamous cell carcinoma proliferation and migration in vitro

Background:Galectin 2(LGALS2)is a protein previously reported to serve as a mediator of disease progression in a range of cancers.The function of LGALS2 in oral squamous cell carcinoma(OSCC),however,has yet to be explored,prompting the present study to address this literature gap.Methods:Overall,144 paired malignant tumor tissues and paracancerous OSCC patient samples were harvested and the LGALS2 expression levels were examined through qPCR and western immunoblotting.The LGALS2 coding sequence was introduced into the pcDNA3.0 vector,to enable the overexpression of this gene,while an LGALS2-specific shRNA and corresponding controls were also obtained.The functionality of LGALS2 as a regulator of the ability of OSCC cells to grow and undergo apoptotic death in vitro was assessed through EdU uptake and CCK-8 assays,and flow cytometer,whereas a Transwell system was used to assess migratory activity and invasivity.An agonist of the Janus Kinase 2(JAK2)/Signal Transducer and Activator of Transcription 3(STAT3)pathway was also used to assess the role of this pathway in the context of LGALS2 signaling.Results:Here,we found that lower LGALS2 protein and mRNA expression were evident in OSCC tumor tissue samples,and these expression levels were associated with clinicopathological characteristics and patient survival outcomes.Silencing LGALS2 enhanced proliferation in OSCC cells while rendering these cells better able to resist apoptosis.The opposite was instead observed after LGALS2 was overexpressed.Mechanistically,the ability of LGALS2 to suppress the progression of OSCC was related to its ability to activate the JAK/STAT3 signaling axis.Conclusion:Those results suggest a role for LGALS2 as a suppressor of OSCC progression through its ability to modulate JAK/STAT3 signaling,supporting the potential utility of LGALS2 as a target for efforts aimed at treating OSCC patients.

Mechanism of Yanghe Pingchaun granules on airway remodeling in asthmatic rats based on IL-6/JAK2/STAT3 signaling axis

Objective: To investigate the effects of Yanghe Pingchuan Granules on airway remodeling in asthmatic rats, and to explore the mechanism of Interleukin-6/Janus kinase 2/ Signal transducing activator of transcription 3(IL-6/JAK2/STAT3) signal axis. Methods: We separated 42 healthy male SD rats into two groups, a control group (7) and a model group (35).The model group was sensitized with a combination of ovalbumin (OVA) and aluminum hydroxide for 2 weeks, while the control group was given an equal amount of physiological saline.After 2 weeks, the modeling group was randomly divided into Model group, Yanghe Pingchuan Granules high, medium and low dose groups and Dexamethasone group, each group consisted of 7 animals. After 4 weeks, OVA atomization and gavage were used for stimulation and treatment. Yanghe Pingchuan Granules high, middle and low groups were given 15.48, 7.74, 3.87 g∙kg-1 Yanghe Pingchuan Granules daily, dexamethasone group was given 0.0625 mg∙kg-1 dexamethasone daily, and the other groups were given the same amount of normal saline. HE, PAS and Masson staining were used to observe the lung histopathological changes in rats. The levels of interleukin-6, IL-23 and IL-17A were detected by ELISA. The expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 in lung tissues were detected by Western blot. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression levels of IL-6, JAK2 and STAT3 in rat lung tissue. Results: The lung tissue structure of the model group was severely damaged compared to the control group, accompanied by a great many of inflammatory cell infiltration, goblet cell hyperplasia, subepithelial collagen fiber deposition and airway epithelial thickening were more obvious. The expressions of IL-6, IL- 23 and IL-17A in serum were significantly increased (P<0.01), the protein expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 and the mRNA expression levels of IL-6, JAK2 and STAT3 in lung tissue were significantly increased (P<0.01);Compared with the model group, inflammatory cell infiltration, goblet cell proliferation, subepithelial collagen fiber deposition and airway epithelial thickening were significantly reduced in each administration group, and the expressions of IL-6, IL-23 and IL-17A in serum were significantly decreased (P< 0.01). The protein expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 and mRNA expression levels of IL-6, JAK2 and STAT3 in lung tissue were significantly decreased (P<0.01). Conclusion: Yanghe Pingchuan Granules can significantly alleviate airway remodeling in asthmatic rats, and its mechanism may be through inhibiting the IL-6/JAK2/STAT3 signal axis.

STAT3 Correlates with Stem Cell-related Transcription Factors in Cervical Cancer

Summary： Cancer stem cells （CSCs） are considered responsible for the high recurrence rate in cervical carcinoma. It has been demonstrated that the signal transducer and activator of transcription 3 （STAT3） is involved in the oncogenesis and takes part in mediating the effects of maintaining stem cell phenotype and pluripotency by regulating the expression of stem cell-related transcription factors. However, the correlation between STAT3 and stem cell-related transcription factors in cervical cancer has not been elucidated. In this study, we established overexpressing plasmid （GV316-STAT3） and siRNA-STAT3 for transfecting Siha cells. Cells negative or positive for Nanog, Oct4, or Sox2 were selected by flow cytometry. Proliferation and differentiation rate of Siha cells was determined by detecting the efficiency of tumor sphere formation. The expression of Nanog, Oct4 and Sox2 （cancer stem cell markers） and STAT3 was detected by quantitative real-time PCR and immunoblotting for Siha cells and by immuno- histochemistry （IHC） for cervical tissues, respectively. The results showed that Nanog＋, Oct4＋, and Sox2＋ Siha-STAT3 over-expressing cells displayed the typical non-adherent spheres. The sphere for- mation efficiency was significantly different between Siha-STAT3 overexpressing cells and siRNA-STAT3 cells （P〈0.05）. Meanwhile, the expression levels of Oct4, Nanog and Sox2 rrtRNA and protein were significantly higher in Siha-STAT3 overexprssing cells than in siRNA-STAT3 cells （P〈0.05）. In addition, the positive rate of STAT3, Nanog, Oct4 and Sox2 in cervical cancer tissues was higher than that in chronic cervicitis group （P〈0.05）. There was a significantly positive relationship between STAT3 and Nanog or Oct4 or Sox2 expression （all P〈0.001）. These results suggested that Oct4＋, Sox2＋, and Nanog＋ cell population possesses stem cell properties in cervical cancer, which may contribute to cervical carcinogenesis and be regulated by STAT3.

Caprylic Acid Improves Lipid Metabolism, Suppresses the Inflammatory Response and Activates the ABCA1/p-JAK2/pSTAT3 Signaling Pathway in C57BL/6J Mice and RAW264.7 Cells

Objective This study aimed to investigate the effects of caprylic acid(C8:0)on lipid metabolism and inflammation,and examine the mechanisms underlying these effects in mice and cells.Methods Fifty-six 6-week-old male C57BL/6J mice were randomly allocated to four groups fed a highfat diet(HFD)without or with 2%C8:0,palmitic acid(C16:0)or eicosapentaenoic acid(EPA).RAW246.7 cells were randomly divided into five groups:normal,lipopolysaccharide(LPS),LPS+C8:0,LPS+EPA and LPS+cAMP.The serum lipid profiles,inflammatory biomolecules,and ABCA1 and JAK2/STAT3 mRNA and protein expression were measured.Results C8:0 decreased TC and LDL-C,and increased the HDL-C/LDL-C ratio after injection of LPS.Without LPS,it decreased TC in mice(P<0.05).Moreover,C8:0 decreased the inflammatory response after LPS treatment in both mice and cells(P<0.05).Mechanistic investigations in C57BL/6J mouse aortas after injection of LPS indicated that C8:0 resulted in higher ABCA1 and JAK2/STAT3 expression than that with HFD,C16:0 and EPA,and resulted in lower TNF-α,NF-κB mRNA expression than that with HFD(P<0.05).In RAW 264.7 cells,C8:0 resulted in lower expression of pNF-κBP65 than that in the LPS group,and higher protein expression of ABCA1,p-JAK2 and p-STAT3 than that in the LPS and LPS+cAMP groups(P<0.05).Conclusion Our studies demonstrated that C8:0 may play an important role in lipid metabolism and the inflammatory response,and the mechanism may be associated with ABCA1 and the p-JAK2/p-STAT3 signaling pathway.

先天性肠闭锁患儿组织STAT3和血清STAT3 mRNA,IL-12p40,IL-13Rα2水平表达及与预后的相关性研究

目的探究先天性肠闭锁(congenital intestinal atresia)患儿组织信号传导与转录激活因子3(signal transducerand activator of transcription 3,STAT3)和血清STAT3 mRNA,IL-12p40及IL-13Rα2水平表达及与预后的相关性。方法收集2020年1月~2023年1月在河北省儿童医院进行治疗的先天性肠闭锁患儿术中切除的100例肠闭锁病变组织、正常肠管组织及术前血清样本,根据Grosfeld分型标准将患儿分为Ⅰ型39例,Ⅱ型22例,Ⅲ型30例和Ⅳ型9例。根据术后6个月的恢复情况,将患儿分为预后良好组(n=78)和预后不良组(n=22);并选取93例同期体检健康儿童的血清样本作为对照。免疫组织化学检测STAT3在组织中阳性表达及定位情况;Western blot检测组织中STAT3蛋白表达;荧光定量PCR法(qPCR)检测血清中STAT3 mRNA的表达水平。采用Pearson相关分析先天性肠闭锁患儿血清STAT3与炎症因子水平的相关性。采用Logistic回归分析影响先天性肠闭锁患儿预后的因素。利用受试者工作特征(ROC)曲线分析血清STAT3水平对先天性肠闭锁患儿预后的预测效能。结果免疫组织化学结果显示,STAT3阳性表达主要定位于细胞质和细胞核,先天性肠闭锁组织中的阳性表达率(86%)显著高于正常肠管组织(18%),差异具有统计学意义(χ^(2)=92.628,P<0.05)。Western blot结果显示STAT3在先天性肠闭锁组织中的相对表达量(1.59±0.21)显著高于正常肠管组织中的相对表达水平(0.81±0.12),差异具有统计学意义(t=30.567,P<0.05)。qPCR法结果显示先天性肠闭锁组患儿血清STAT3 mRNA(2.13±0.56),IL-12p40(0.89±0.13ng/ml)以及IL-13Rα2(6.42±1.86ng/ml)水平均显著高于对照组(1.06±0.11,0.37±0.08ng/ml,1.35±0.41ng/ml),差异具有统计学意义(t=18.101,33.170,25.708,均P<0.05)。随着患儿分型的增加,STAT3 mRNA及IL-12p40,IL-13Rα2水平逐渐增加,差异具有统计学意义(F=52.666,160.300,25.82,均P<0.05)。Pearson相关分析显示先天性肠闭锁患儿血清STAT3 mRNA与炎症因子IL-12p40,IL-13Rα2水平均呈显著正相关(r=0.496,0.564,均P<0.001)。先天性肠闭锁患儿预后不良组血清STAT3 mRNA(3.01±0.75)表达水平显著高于预后良好组(1.88±0.51),差异具有统计学意义(t=8.212,P<0.05)。Logistic回归分析结果显示,STAT3 mRNA,IL-12p40,IL-13Rα2水平以及低出生质量均是先天性肠闭锁患儿预后不良的独立危险因素(均P<0.05)。ROC曲线可知血清STAT3评估先天性肠闭锁患儿预后的曲线下面积(AUC)为0.916,敏感度和特异度分别为81.82%,88.46%,当血清STAT3 mRNA水平高于2.47时先天性肠闭锁患儿发生预后不良的几率较高。结论STAT3在先天性肠闭锁患儿组织和血清中的表达显著升高,血清STAT3对患儿预后情况具有一定的预测价值。

miR-125a-5p通过靶向STAT3减轻戊四唑诱导的癫痫大鼠神经功能障碍和炎症反应

目的:探讨miR-125a-5p通过靶向信号转导与转录激活因子3(STAT3)对戊四唑(PTZ)诱导的癫痫大鼠神经功能障碍和炎症反应的影响。方法:采用PTZ点燃法建立大鼠癫痫模型,并将其随机分为6组:Con组、PTZ组、PTZ+agomir NC组、PTZ+miR-125a-5p agomir组、PTZ+miR-125a-5p agomir+pcDNA组和PTZ+miR-125a-5p agomir+pc-STAT3组,评价miR-125a-5p对大鼠癫痫持续时间、潜伏期、惊厥次数和严重程度的影响。qRT-PCR检测miR-125a-5p和STAT3 mRNA表达;改良神经系统严重程度评分(mNSS)、开场实验和Rotarod测试评价癫痫大鼠的神经功能;Morris水迷宫实验评价癫痫大鼠的认知功能;免疫染色观察海马CA1和CA3区域神经元凋亡;ELISA检测海马组织炎症因子(TNF-α、IL-6、IL-1β和IL-18)水平;Western blot检测海马组织中STAT3蛋白表达;双荧光素酶实验验证miR-125a-5p和STAT3的关系。结果:在PTZ诱导的癫痫大鼠中,miR-125a-5p表达上调,STAT3表达下调,且miR-125a-5p靶向抑制STAT3表达。与PTZ组相比,PTZ+miR-125a-5p agomir组癫痫大鼠的潜伏期缩短,癫痫评分降低,癫痫发作时间和次数减少,mNSS降低,掉落潜伏期及外围区域的移动距离、开放区域的总距离延长,逃避潜伏期显著降低,穿越平台次数及停留在目标象限的时间和游泳距离均增加,海马组织TNF-α、IL-6、IL-1β、IL-18表达降低,CA3和CA1区的NenN+细胞增多(P<0.05);STAT3过表达可逆转miR-125a-5p agomir对癫痫大鼠神经功能障碍及炎症反应的缓解作用(P<0.05)。结论:miR-125a-5p通过靶向下调STAT3,减轻PTZ诱导的癫痫大鼠神经功能障碍和炎症反应。

冬凌草甲素调节JAK2/STAT3/SOCS-1信号通路对糖耐量异常大鼠胰岛素抵抗的影响

目的探讨冬凌草甲素(Oridonin,Ori)对糖耐量异常(impaired glucose tolerance,IGT)大鼠胰岛素抵抗(insulin resistance,IR)的影响及作用机制。方法采用高脂饮食喂养联合链脲佐菌素注射法构建IGT大鼠IR模型,大鼠分为正常组(CT组)、IGT模型组(IGT组)、Ori组(10 mg·kg^(-1)·d^(-1))、Ori+Colivelin(COL)组(10 mg·kg^(-1)·d^(-1)Ori+2 mg/kg COL),每组6只。血糖检测仪测定空腹血糖(FPG)、葡萄糖耐量试验(OGTT)2 h血糖(2 hPG),ELISA试剂盒测定空腹胰岛素(FINS)、单核细胞趋化蛋白1(MCP-1)、肿瘤坏死因子α(TNF-α)含量,计算胰岛素抵抗指数(HOMA-IR),血液自动分析仪测定血清胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)水平,HE染色观察肝脏病理形态,Western blot验证附睾脂肪磷酸化(p)-激活Janus激活激酶2(JAK2)、JAK2、p-信号转导和转录激活因子3(STAT3)、STAT3、p-细胞因子信号传导抑制蛋白1(SOCS-1)、SOCS-1蛋白表达。结果与CT组比较,IGT组大鼠肝脏细胞肿胀,胞浆内可见大量大小不一的脂肪空泡,细胞核被脂肪空泡挤压偏位,且发生炎性细胞浸润,FPG、2hPG、FINS、HOMA-IR、TC、TG、LDL-C、MCP-1、TNF-α以及p-JAK2/JAK2、p-STAT3/STAT3、p-SOCS-1/SOCS-1蛋白表达水平升高,血清HDL-C水平下降(P<0.05);与IGT组相比,Ori组大鼠肝脏细胞胞浆内脂肪滴及空泡数量明显减少,细胞肿胀有所缓解,未见炎性细胞浸润,FPG、2hPG、FINS、HOMA-IR、TC、TG、LDL-C、MCP-1、TNF-α以及p-JAK2/JAK2、p-STAT3/STAT3、p-SOCS-1/SOCS-1蛋白表达水平下降,血清HDL-C水平升高(P<0.05);与Ori组相比,Ori+COL组大鼠肝脏脂肪变状况加剧,细胞肿大,血清FPG、2hPG、FINS、HOMA-IR、TC、TG、LDL-C、MCP-1、TNF-α以及p-JAK2/JAK2、p-STAT3/STAT3、p-SOCS-1/SOCS-1蛋白表达水平升高,血清HDL-C水平下降(P<0.05)。结论Ori对IGT大鼠IR的缓解作用可能与抑制JAK2/STAT3/SOCS-1信号通路激活有关。

Glycine Attenuates Myocardial Fibrosis in Myocardial Infarction in Rats Partly through Modulating Signal Transducer and Activator of Transcription 3/Nuclear Factor-κB/Transforming Growth Factor-β axis

Objective: Inflammation and fibrosis are strongly associated with each other. Glycine is present in various traditional Chinese medicines and exhibits anti-inflammatory activity. However, the effects of glycine on myocardial fibrosis(MF) in rats with myocardial infarction(MI) have not been reported. The purpose of this study is to investigate the effects of glycine therapy on MF and comprehend its underlying mechanisms. Materials and Methods: Left anterior descending artery ligation-induced MI in Sprague Dawley rats was leveraged to assess the therapeutic effects of Glycine. Rats received either normal saline or glycine(0.5 mg/g bodyweight) for 7 days. Results: Glycine upregulated cardiac ejection fraction and fractional shortening to improve cardiac function, as evaluated by echocardiography. Histological and immunohistochemical analyses demonstrated that glycine could decrease inflammatory cell infiltration and alleviate collagen deposition. Western blotting revealed that nuclear factor-κB(NF-κB)-mediated inflammatory signaling was also downregulated by glycine treatment. The expression of signal transducer and activator of transcription 3(STAT3), tumor necrosis factor-α, and transforming growth factor-β(TGF-β) was decreased significantly in the glycine-treated group compared to the model group. Thus, glycine plays a protective role against myocardial ischemia and subsequent MF. Conclusion: The protective effects of glycine were achieved partly through STAT3/NF-κB/TGF-β signaling pathway.

川崎病合并支原体感染病儿瓣膜炎性损伤与IL-6/STAT3信号通路表达变化的关系分析

目的探讨川崎病(KD)合并支原体感染病儿瓣膜炎性损伤与白细胞介素-6(IL-6)/信号转导及转录激活蛋白3(STAT3)信号通路表达变化的关系。方法前瞻性选取2018年1月至2022年12月安阳市妇幼保健院收治的50例KD合并肺炎支原体(MP)(KD-MP)感染病儿为感染组,另选取同期收治的106例单纯KD病儿为未感染组,比较感染组与未感染组病儿瓣膜炎性损伤情况。再根据KD病儿是否伴有冠状动脉损伤(CAL)损伤分为CAL组(n=84)、无CAL组(n=72),比较两组病儿IL-6和STAT3表达水平,通过多因素logistic回归分析KD病儿发生CAL的危险因素。比较KD并CAL不同预后病儿IL-6和STAT3表达变化,使用受试者操作特征曲线(ROC曲线)分析IL-6、STAT3水平对KD并CAL病儿预后的预测价值。结果感染组KD病儿发热时间、住院时间、初诊CAL发生率均高于未感染组(P<0.05);感染组KD病儿C反应蛋白(CRP)、降钙素原(PCT)、红细胞沉降率(ESR)、IL-6 mRNA表达水平和STAT3 mRNA表达水平均高于未感染组[(51.32±12.62)mg/L比(36.58±10.98)mg/L,(1.33±0.35)μg/L比(1.21±0.31)μg/L,(84.69±15.66)mm/h比(49.62±16.43)mm/h,4.14±1.09比3.06±1.15,2.81±0.89比1.68±0.54,均P<0.05]。CAL组病儿IL-6、STAT3 mRNA表达水平高于无CAL组(4.13±1.22比2.56±0.87,2.46±0.71比1.55±0.44,均P<0.05);多因素回归分析结果显示,MP感染、IL-6 mRNA、STAT3 mRNA表达是KD病儿发生CAL的独立危险因素(均P<0.05)。KD并CAL预后良好组病儿IL-6、STAT3 mRNA表达水平低于预后不良组(3.57±1.06比4.57±1.13,1.94±0.58比2.87±0.87,均P<0.05),ROC曲线结果显示IL-6、STAT3及其联合预测KD并CAL预后的曲线下面积(AUC)分别为0.75、0.81、0.91,且联合检测的AUC高于IL-6、STAT3单独检测(均P<0.05)。结论KD合并MP病儿瓣膜炎性损伤更为严重,IL-6/STAT3信号通路活化是其危险因素,且该信号通路对KD并CAL病儿预后有较高的预测价值。

嘎木珠尔灌肠通过对IL-6/STAT3信号通路的调节而改善结肠炎模型大鼠的结肠炎症

目的:观察嘎木珠尔灌肠对结肠炎模型大鼠结肠病变的影响;并研究其对IL-6/STAT3信号通路的调节作用。方法:40只大鼠被随机分组;空白组、造模组、柳氮磺吡啶组及嘎木珠尔组,每组10只。以乙醇/TNBS灌肠造模后分别以生理盐水、柳氮磺吡啶及嘎木珠尔灌肠。14天后对结肠大体形态及结肠病理切片进行观察,免疫组化分析结肠IL-6及STAT3的表达。结果:造模后结肠大体评分及组织病理评分均显著增高(P均<0.01);结肠IL-6及STAT3阳性染色细胞数量均显著增多(P均<0.01)。经柳氮磺吡啶及嘎木珠尔治疗后结肠大体评分均明显降低(P<0.01、P<0.05);结肠组织病理评分均显著降低(P均<0.01);结肠IL-6及STAT3阳性染色细胞数均显著减少(P均<0.01)。结论:嘎木珠尔灌肠能减轻结肠炎模型大鼠的结肠炎症,并可改善其病理结构的破坏。其可能通过对IL-6/STAT3信号通路下调而发挥作用。