Exploring the mechanism of electroacupuncture at different acupoints on acute colitis rats based on JAK2/STAT3/SOCS1 signaling pathway

Objective:To investigate the mechanism of JAK2/STAT3/SOCS1 signaling pathway in electroacupuncture of different acupoints on acute colitis rats.Methods:36 SPF SD rats were randomly divided into 6 groups,with 6 rats in each group.The rat model of acute colitis was prepared by enema with glacial acetic acid solution.After the model was established,electroacupuncture was given to each acupoint group,with density wave,frequency 2Hz-50 Hz,intensity 2 mA,muscle tremor as the degree 20 min/time,1 time/day,for 3 consecutive days.Observe the general condition of rats;the pathological changes of colonic mucosa in rats were observed by HE method.The contents of serum interleukin-4(IL-4)and interleukin-8(IL-8)were detected by ELISA.Western blot and RT-PCR were used to detect the expression of JAK2,STAT3,SOCS1 protein and mRNA in rat colon tissue.Results:In contrast to the normal group,the overall condition of the model group was worse,the colonic mucosa was severely damaged,even necrotic,and the ulcer surface was obvious.The content of IL-4 in serum was obviously reduced,and the content of IL-8 was obviously go up(P<0.01).The protein content of JAK2,STAT3 and the expression of JAK2,STAT3 mRNA in colon tissue of rats were obviously go up,while the protein content of SOCS1 and the expression of SOCS1 mRNA were obviously reduced(P<0.01).In contrast to the model group,the general condition of rats in each acupoint group was significantly improved,the damage and necrosis of colonic mucosa and ulcer surface were obviously alleviated,the content of IL-4 in serum was obviously go up,and the content of IL-8 was significantly decreased(P<0.01).The protein content of JAK2,STAT3 and the expression of JAK2,STAT3 mRNA in colon tissue of rats were obviously reduced,while the protein content of SOCS1 and the expression of SOCS1 mRNA were obviously go up(P<0.05,P<0.01).Comparison of different acupoint groups,the colonic mucosal injury in the Zusanli group was significantly reduced,the content of serum IL-4 was significantly increased,and the content of IL-8 was significantly decreased(P<0.05,P<0.01).The protein content and mRNA expression of JAK2 and STAT3 in colon tissue were significantly down-regulated,while the protein content and mRNA expression of SOCS1 were significantly go up(P<0.05,P<0.01).Conclusion:Electroacupuncture at each acupoint can improve the damage of colonic mucosa and reduce the inflammatory response.The therapeutic effect of Zusanli(ST36)is better than that of Tianshu(ST25),Dachangshu(BL25)and Shangjuxu(ST37).The mechanism may be related to the regulation of JAK2/STAT3/SOCS1 signaling pathway related proteins and inflammatory cytokines IL-4 and IL-8.

Yangyin Huowei mixture alleviates chronic atrophic gastritis by inhibiting the IL-10/JAK1/STAT3 pathway

BACKGROUND The Chinese medicine Yangyin Huowei mixture(YYHWM)exhibits good clinical efficacy in the treatment of chronic atrophic gastritis(CAG),but the mechanisms underlying its activity remain unclear.AIM To investigate the therapeutic effects of YYHWM and its underlying mechanisms in a CAG rat model.METHODS Sprague-Dawley rats were allocated into control,model,vitacoenzyme,and low,medium,and high-dose YYHWM groups.CAG was induced in rats using Nmethyl-N′-nitro-N-nitrosoguanidine,ranitidine hydrochloride,hunger and satiety perturbation,and ethanol gavage.Following an 8-wk intervention period,stomach samples were taken,stained,and examined for histopathological changes.ELISA was utilized to quantify serum levels of PG-I,PG-II,G-17,IL-1β,IL-6,and TNF-α.Western blot analysis was performed to evaluate protein expression of IL-10,JAK1,and STAT3.RESULTS The model group showed gastric mucosal layer disruption and inflammatory cell infiltration.Compared with the blank control group,serum levels of PGI,PGII,and G-17 in the model group were significantly reduced(82.41±3.53 vs 38.52±1.71,23.06±0.96 vs 11.06±0.70,and 493.09±12.17 vs 225.52±17.44,P<0.01 for all),whereas those of IL-1β,IL-6,and TNF-αwere significantly increased(30.15±3.07 vs 80.98±4.47,69.05±12.72 vs 110.85±6.68,and 209.24±11.62 vs 313.37±36.77,P<0.01 for all),and the protein levels of IL-10,JAK1,and STAT3 were higher in gastric mucosal tissues(0.47±0.10 vs 1.11±0.09,0.49±0.05 vs 0.99±0.07,and 0.24±0.05 vs 1.04±0.14,P<0.01 for all).Compared with the model group,high-dose YYHWM treatment significantly improved the gastric mucosal tissue damage,increased the levels of PGI,PGII,and G-17(38.52±1.71 vs 50.41±3.53,11.06±0.70 vs 15.33±1.24,and 225.52±17.44 vs 329.22±29.11,P<0.01 for all),decreased the levels of IL-1β,IL-6,and TNF-α(80.98±4.47 vs 61.56±4.02,110.85±6.68 vs 89.20±8.48,and 313.37±36.77 vs 267.30±9.31,P<0.01 for all),and evidently decreased the protein levels of IL-10 and STAT3 in gastric mucosal tissues(1.11±0.09 vs 0.19±0.07 and 1.04±0.14 vs 0.55±0.09,P<0.01 for both).CONCLUSION YYHWM reduces the release of inflammatory factors by inhibiting the IL-10/JAK1/STAT3 pathway,alleviating gastric mucosal damage,and enhancing gastric secretory function,thereby ameliorating CAG development and cancer transformation.

Oleanolic acid inhibits colon cancer cell stemness and reverses chemoresistance by suppressing JAK2/STAT3 signaling pathway

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.Octamer-binding transcription factor 4(Oct4),Prominin-1(CD133),Nanog,and transcription factor SOX-2(SOX2)are cell stemness-related indicators whose expression was assessed usingfluorescence 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:Thesefindings 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.

Effects of plumbagin on migration and invasion of human hepatoma cell line via JAK2/STAT3 signaling pathway

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.

Morroniside ameliorates lipopolysaccharide-induced inflammatory damage in iris pigment epithelial cells through inhibition of TLR4/JAK2/STAT3 pathway

AIM:To investigate the effect of morroniside(Mor)on lipopolysaccharide(LPS)-treated iris pigment epithelial cells(IPE).METHODS:IPE cells were induced by LPS and treated with Mor.Cell proliferation was detected by cell counting kit(CCK)-8,apoptosis was detected by flow cytometry,the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-8 were measured by enzyme-linked immunosorbent assay(ELISA)kits,and the protein expression of TLR4,JAK2,p-JAK2,STAT3,and p-STAT3 was analyzed by Western blotting.In addition,overexpression of TLR4 and Mor treatment of LPS-stimulated IPE cells were also tested for the above indices.RESULTS:Mor effectively promoted the proliferation and inhibited the apoptosis of LPS-treated IPE cells.In addition,Mor significantly reduced the levels of TNF-α,IL-6,and IL-8 and significantly inhibited the expression of TLR4,p-JAK2,and p-STAT3 in LPS-treated IPE cells.The effect of Mor on LPS-treated IPE cells was markedly attenuated after overexpression of TLR4.CONCLUSION:These findings suggest that Mor may ameliorate LPS-induced inflammatory damage and apoptosis in IPE through inhibition of TLR4/JAK2/STAT3 pathway.

To explore the mechanism of Dahuang Lingxian Formula in relieving inflammatory response of bile duct cells based on IL-6/JAK/STAT3 signaling pathway

Objective:To explore the mechanism of action of Dahuang Lingxian Formula in alleviating the inflammatory response of bile duct cells in LPS-induced intrahepatic bile duct inflammation model rats based on IL-6/JAK/STAT3 signaling pathway.Methods:Fifty SD rats were randomly divided into five groups,blank group,model group,choling tablets(0.5 g/kg),and low and high concentration groups(2.4 g/kg and 4.8 g/kg)of Dahuang Lingxian Formula,ten rats in each group.Except for the blank group,the rats in each group were injected with 1.25 mg/kg LPS at the common bile duct at one time to construct an animal model of intrahepatic bile duct infection.After gavage on day 8,liver tissues were taken from rats at the hepatic hilum,and the histopathological changes of the hepatic hilum and biliary tree were observed by HE staining.The expression levels of serum glutamic alanine transaminase(ALT),glutamic oxalacetic transaminase(AST),malondialdehyde(MDA)and superoxide dismutase(SOD)were measured by biochemical method.The expression levels of interleukin 6(IL-6),Janus protein tyrosine kinase 2(JAK2),signal transducer and activator of transcription 3(STAT3)in rat serum were measured by enzyme-linked immunosorbent assay(ELISA).Protein immunoblotting(WB)and real-time fluorescence quantitative PCR(RT-qPCR)were used to detect the expression levels of IL-6,JAK2,STAT3 protein and mRNA in biliary tree tissues.Results:①Compared with the blank group,the structures such as interlobular bile ducts in the hepatic sinusoids and portal duct area of the model rats were destroyed,and inflammatory cells infiltrated around them.The expression of ALT,AST,MDA,IL-6,JAK2 and STAT3 in the serum increased significantly,the expression level of SOD decreased,and the expression levels of IL-6,JAK2 and STAT3 proteins and mRNA increased.②Compared with the model group,the degree of liver pathological damage in rats in the Chiling Ning tablet group and the low and high concentration groups of Dahuang Lingxian Formula were improved,which could significantly reduce the expression levels of ALT,AST,MDA,IL-6,JAK2,STAT3 and up-regulate SOD in serum,and down-regulate the expression of IL-6,JAK2,STAT3 protein and mRNA,with the best effect in the high concentration group of Dahuang Lingxian Formula.③Compared with the choling tablet group,the rats in the low and high concentration groups of Dahuang Lingxian Formula tended to normalize the degree of liver pathological damage,without obvious inflammatory cell infiltration,and the expression levels of ALT,AST,MDA,IL-6,JAK2,STAT3 and the expression levels of IL-6,JAK2,STAT3 protein and mRNA in serum were reduced,and the expression levels of SOD were increased,with the best effect of Dahuang Lingxian Formula The treatment effect was best in the high concentration group.Conclusion:The mechanism may be related to the down-regulation of IL-6/JAK/STAT3 signaling pathway activation,and the best therapeutic effect was achieved by the high concentration group of Dahuang Lingxian Formula.

水杨酸通过AMPK/STAT3改善同型半胱氨酸诱导的大鼠学习记忆障碍和炎症反应

目的探讨水杨酸(salicylic acid,SA)对同型半胱氨酸(homocysteine,Hcy)诱导的阿尔茨海默症(Alzheimer's disease,AD)大鼠模型认知障碍的改善作用及其可能机制。方法将36只健康雄性SD大鼠随机分为对照组、模型组和治疗组三组,每组各12只。模型组通过连续14天尾静脉注射Hcy(400μg/kg/d)构建AD模型,治疗组造模方法同模型组。治疗组大鼠给予水杨酸灌胃治疗,持续4周;对照组和模型组每天给予同等体积的生理盐水灌胃,持续4周。通过Morris水迷宫检测各组大鼠学习记忆能力;ELISA法检测各组大鼠血清中炎症因子IL-1β和TNF-α的分泌情况;Western bolt法检测各组大鼠额叶皮质腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)、磷酸化AMPK(p-AMPK)和信号传导及转录激活因子3(signal transducer and activator of transcription 3,STAT3)蛋白的表达情况;免疫组织化学染色方法检测各组大鼠额叶皮质内p-AMPK、STAT3表达情况。结果与对照组相比,模型组大鼠逃避潜伏期显著延长(P<0.001),穿越原平台次数和平台象限滞留时间百分比均显著减少(P<0.001),炎症因子IL-1β和TNF-α分泌增多(P<0.01),额叶组织p-AMPK蛋白表达降低(P<0.01),STAT3蛋白表达增高(P<0.001);与模型组相比,治疗组大鼠逃避潜伏期显著缩短(P<0.05),穿越原平台次数和平台象限滞留时间百分比均显著增多(P<0.05),炎症因子IL-1β和TNF-α分泌减少(P<0.05),额叶组织p-AMPK蛋白表达显著增高(P<0.05),STAT3蛋白表达显著降低(P<0.05)。结论水杨酸在一定程度上改善了Hcy诱导的AD大鼠学习记忆能力,其机制可能与激活AMPK抑制STAT3蛋白表达有关。

CaMKKβ通过激活AMPK/JAK2/STAT3信号促进小鼠单核巨噬细胞向M2表型转换

目的·探明钙离子/钙调蛋白依赖性蛋白激酶激酶β(CaMKKβ)在白介素4(IL-4)诱导的RAW264.7细胞极化过程中的作用和机制。方法·RT-PCR和ELISA检测IL-4诱导RAW264.7细胞极化水平,Western blotting测定极化过程中CaMKKβ、AMPK、JAK2、STAT3蛋白表达及磷酸化水平。慢病毒为载体的shRNA稳定干扰RAW264.7细胞CaMKKβ表达,RT-PCR和ELISA检测IL-4诱导下细胞极化水平的改变,Western blotting检测AMPK、JAK2、STAT3蛋白及磷酸化蛋白的表达。分别特异性阻断AMPK、JAK2和STAT3蛋白活性,经RT-PCR检测IL-4诱导下RAW264.7细胞极化水平的改变。结果·IL-4主要诱导RAW264.7细胞向M2型巨噬细胞极化(P<0.05),且CaMKKβ、AMPK、JAK2、STAT3的磷酸化水平明显升高(均P<0.05)。shRNA稳定干扰RAW264.7细胞CaMKKβ表达后,IL-4促进巨噬细胞向M2型极化作用降低(P<0.05),且AMPK、JAK2和STAT3的磷酸化水平降低(均P<0.05)。分别阻断AMPK、JAK2和STAT3,巨噬细胞向M2型极化减少(均P<0.05)。结论·CaMKKβ通过激活AMPK/JAK2/STAT3信号通路,促进IL-4诱导的巨噬细胞M2型极化。

脂联素导致急性心力衰竭大鼠AMPK STAT3蛋白及血清炎性因子含量的变化

目的探讨脂联素(adiponectin,ADPN)对急性心力衰竭(AHF)大鼠AMP活化蛋白激酶(AMP-activated protein kinase,AMPK)、信号转导子与转录激活子3(signal transducer and activator of transcription 3,STAT3)通路相关蛋白表达变化和血清炎性因子含量变化的影响。方法将30只雄性SD大鼠随机分为对照组(CON组)、模型组(AHF组)和脂联素处理组(AHFADPN组),AHF-ADPN组大鼠尾静脉注射重组脂联素腺病毒7 d(10×10^(9)pfu,1天1次),AHF组大鼠注射等剂量的生理盐水。AHF组和AHF-ADPN组大鼠通过异丙肾上腺素(isoprenaline,ISO)腹腔注射构建AHF模型。酶联免疫吸附测定方法检测三组大鼠血清B型尿钠肽(B-type natriuretic peptides,BNP)、心肌肌钙蛋白I (cardiac troponin I, cTnI)、乳酸脱氢酶(lactate dehydrogenase,LDH)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-10(interleukin-10,IL-10)和生长分化因子-15(growthdifferentiation factor-15,GDF-15)含量;高分辨率小动物超声系统比较三组大鼠心率(heart rate,HR)及左室射血分数(left ventricular ejection fraction,LVEF);免疫印迹方法检测三组大鼠pAMPK、AMPK、p-STAT3和STAT3蛋白表达变化。结果①与AHF组比较,AHF-ADPN组血清BNP含量[(286.30±14.03) pg/mL vs.(79.95±0.85) pg/mL]显著下调(P <0.0001),cTnI含量[(543.80±17.30) pg/mL vs.(147.40±21.39) pg/mL]显著下调(P <0.001),LDH含量[(6.91±0.07) U/L vs.(6.37±0.08) U/L]显著下调(P <0.01)。②与AHF组比较,AHF-ADPN组HR[(335.40±4.67)次/min vs.(387.80±3.74)次/min]显著升高(P <0.001),LVEF[(42.10±2.76)%vs.(79.20±1.93)%]显著升高(P <0.001)。③与AHF组比较,AHF-ADPN组血清IL-6含量[(280.00±4.63) pg/mL vs.(251.70±5.59) pg/mL]显著降低(P <0.01),TNF-α含量[(87.35±2.09) pg/mL vs.(74.42±2.51) pg/mL]显著降低(P <0.001),GDF-15含量[(123.40±3.03)pg/mL vs.(97.69±2.52) pg/mL]显著降低(P <0.001),血清IL-10含量[(101.90±2.46)pg/mL vs.(189.90±4.18) pg/mL]显著升高(P <0.0001)。④AHF-ADPN组p-AMPK及pSTAT3蛋白表达显著高于AHF组(P <0.01)。结论外源ADPN能够显著改善AHF大鼠的血清炎性因子含量,并导致心肌p-AMPK和p-STAT3表达改变,为AHF的治疗提供证据。

基于AMPK/PPARα/STAT3途径探讨姜黄素对单核细胞迁移和血管壁巨噬细胞形成的影响

目的基于腺苷酸活化蛋白激酶/过氧化物酶体增殖物激活受体α/信号传导及转录激活因子3(AMPK/PPARα/STAT3)途径探讨姜黄素对单核细胞迁移和血管壁巨噬细胞形成的影响。方法将人单核细胞株THP-1细胞分为空白对照组、LPS组(lipopolysaccharide,LPS,5μg·mL-1)、LPS+辛伐他汀组(LPS 5μg·mL-1+Simvastatin 5μmol·L-1)、LPS+姜黄素组[用姜黄素(10、20、40、80μmol·L-1)处理2 h后,再用5μg·mL-1LPS刺激THP-1细胞0.5 h];采用Transwell迁移实验检测THP-1细胞迁移能力;采用ELISA法检测高迁移率族蛋白B1(HMGB1)、单核细胞趋化蛋白-1(MCP-1)的蛋白水平;采用RT-PCR法检测MCP-1的mRNA表达;采用Western Blot法检测腺苷酸活化蛋白激酶(AMPK)、过氧化物酶体增殖物激活受体α(PPARα)、信号转导及转录激活因子3(STAT3)的蛋白表达;采用RT-PCR法检测AMPK、PPARα、STAT3的mRNA表达;采用CD68免疫组织化学染色法计算载脂蛋白E(ApoE)小鼠血管壁巨噬细胞的含量。结果与空白对照组相比,LPS组THP-1细胞迁移能力显著升高,HMGB1、MCP-1蛋白及mRNA表达水平显著升高,AMPK、PPARα蛋白及mRNA表达水平均显著降低,STAT3蛋白及mRNA表达水平显著升高(P<0.05或P<0.01);与空白对照组相比,LPS组小鼠血管壁巨噬细胞数量显著升高(P<0.05或P<0.01);与LPS组相比,LPS+姜黄素(10、20、40、80μmol·L-1)组THP-1细胞迁移能力显著降低,HMGB1、MCP-1蛋白及mRNA表达水平显著降低,AMPK、PPARα蛋白及mRNA表达水平均显著升高,STAT3蛋白及mRNA表达水平显著降低(P<0.05或P<0.01)。与LPS组相比,姜黄素各剂量组10、20、40、80 mg·kg-1小鼠血管壁巨噬细胞数量降低(P<0.05或P<0.01);其中高剂量组效果优于低剂量组(P<0.05或P<0.01)。结论姜黄素可抑制LPS诱导的THP-1单核细胞的迁移及血管壁巨噬细胞的形成,其作用机制可能是通过调节AMPK/PPARα/STAT3途径来实现的。

Circular RNA ciRS-7 promotes the proliferation and metastasis of pancreatic cancer by regulating miR-7-mediated EGFR/STAT3 signaling pathway

Background: Pancreatic ductal adenocarcinoma(PDAC) is the most deadly type of tumor, and its pathogenesis remains unknown. Circular RNAs(circRNAs) may be functional and bind to micro RNAs and consequently, influence the activity of targeted mRNAs. Recent researches indicate that one circRNA, ciRS-7, acts as a sponge of miR-7 and thus, inhibits its activity. It is well known that miR-7 is a cancer suppressor in many cancers. However, the relationship between ciRS-7 and miR-7, and the role of ciRS-7 in PDAC, remains to be elucidated. Methods: miR-7 and ciRS-7 expression in 41 pairs of PDAC tumors and their paracancerous tissues were detected by quantitative reverse transcription polymerase chain reaction(qRT-PCR). The relationships between their expression levels and clinicopathological features in PDAC tissues were assessed. The relationship between miR-7 and ciRS-7 was also assessed by Spearman’s correlation. We also used cell lines to evaluate the role of ciRS-7 in cell line behavior. The ciRS-7 interfere RNA(si RNA) and its empty vector were transfected into PDAC cells. PDAC cells proliferation and invasion abilities were detected by MTT assay and invasion analysis. The expression of proteins was assessed by Western blotting. Results: ciRS-7 expression was significantly higher in PDAC tissues than paracancerous tissues( P = 0.002). However, miR-7 expression showed the opposite trend( P = 0.048). Moreover, ciRS-7 expression was inversely correlated with miR-7 in PDAC( r s =-0.353, P = 0.023). ciRS-7 expression was also significantly elevated in venous invasion(3.72 ± 2.93 vs. 2.14 ± 1.26;P = 0.028) and lymph node metastasis(4.19 ± 2.75 vs. 2.32 ± 1.90;P = 0.016) in PDAC patients. Furthermore, ciRS-7 knockdown suppressed cell proliferation and invasion of PDAC cells( P < 0.05), and the downregulation of ciRS-7 resulted in miR-7 overexpression and subsequent inhibition of epidermal growth factor receptor(EGFR) and signal transducer and activator of transcription 3(STAT3). Conclusions: Circular RNA ciRS-7 plays an oncogene role in PDAC, partly by targeting miR-7 and regulating the EGFR/STAT3 signaling pathway.

Elevated retinol binding protein 4 levels are associated with atherosclerosis in diabetic rats via JAK2/STAT3 signaling pathway

BACKGROUND Atherosclerosis is a major cause of mortality worldwide and is driven by multiple risk factors,including diabetes,which results in an increased atherosclerotic burden,but the precise mechanisms for the occurrence and development of diabetic atheroscerosis have not been fully elucidated.AIM To summarize the potential role of retinol binding protein 4(RBP4) in the pathogenesis of diabetic atheroscerosis,particularly in relation to the RBP4-Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway.METHODS Male Wistar rats were randomly divided into three groups,including a control group(NC group),diabetic rat group(DM group),and diabetic atherosclerotic rat group(DA group).The contents of total cholesterol(TC), high-density lipoprotein cholesterol(HDL-c), triglycerides(TG), low-density lipoprotein cholesterol(LDLc), fasting insulin(FINS),fasting plasma glucose,and hemoglobin A1 c(HbA1 c)were measured.Moreover,the adipose and serum levels of RBP4,along with the expression levels of JAK2, phosphorylated JAK2(p-JAK2), STAT3,phosphorylated STAT3(p-STAT3), B-cell lymphoma-2(Bcl-2), and Cyclin D1 in aortic tissues were also measured.Besides,homeostasis model assessment of insulin resistance(HOMA-IR) and atherogenic indexes(AI) were calculated.RESULTS Compared with the NC and DM groups,the levels LDL-c,TG,TC,FINS,HOMAIR,RBP4,and AI were upregulated,whereas that of HDL-c was downregulated in the DA group(P <0.05);the mRNA levels of JAK2,STAT3,Cyclin D1,and Bcl-2 in the DA group were significantly increased compared with the NC group and the DM group;P-JAK2,p-JAK2/JAK2 ratio,p-STAT3,p-STAT3/STAT3 ratio,Cyclin D1,and Bcl-2 at protein levels were significantly upregulated in the DA group compared with the NC group and DM group.In addition,as shown by Pearson analysis,serum RBP4 had a positive correlation with TG,TC,LDL-c,FINS,HbA1 C,p-JAK2,p-STAT3,Bcl-2,Cyclin D1,AI,and HOMA-IR but a negative correlation with HDL-c.In addition,multivariable logistic regression analysis showed that serum RBP4,p-JAK2,p-STAT3,and LDL-c were predictors of the presence of diabetic atherosclerosis.CONCLUSION RBP4 could be involved in the initiation or progression of diabetic atherosclerosis by regulating the JAK2/STAT3 signaling pathway.

3-epi-bufotalin suppresses the proliferation in colorectal cancer cells through the inhibition of the JAK1/STAT3 signaling pathway

Traditional Chinese medicine(TCM)has been increasingly employed in the last decades in China for both preventing and treating a variety of cancers.3-epi-bufotalin is an active ingredient of TCM“Chanpi”with anti-tumor potential.However,the effect and mechanism of 3-epi-bufotalin on colorectal cancers were not well disclosed.The present study demonstrated that 3-epi-bufotalin could reduce viability,trigger apoptosis,and block the cell cycle at the G2/M stage in colorectal cancer cell lines HT29,RKO,and COLO205 in vitro.Moreover,3-epi-bufotalin inhibited the JAK1/STAT3 signaling pathway.These results indicated the anti-proliferation ability of 3-epi-bufotalin in colorectal cancer cells.

Hepatocellular carcinoma-derived exosomal miRNA-761 regulates the tumor microenvironment by targeting the SOCS2/JAK2/STAT3 pathway

BACKGROUND:Exosomes and exosomal microRNAs have been implicated in tumor occurrence and metastasis.Our previous study showed that microRNA-761(miR-761)is overexpressed in hepatocellular carcinoma(HCC)tissues and that its inhibition affects mitochondrial function and inhibits HCC metastasis.The mechanism by which exosomal miR-761 modulates the tumor microenvironment has not been elucidated.METHODS:Exosomal miR-761 was detected in six cell lines.Cell counting kit-8(CCK-8)and transwell migration assays were performed to determine the function of exosomal miR-761 in HCC cells.The luciferase reporter assay was used to analyze miR-761 target genes in normal fi broblasts(NFs).The inhibitors AZD1480 and C188-9 were employed to determine the role of the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway in the transformation of cancer-associated fi broblasts(CAFs).RESULTS:In this study,we characterized the mechanism by which miR-761 reprogrammed the tumor microenvironment.We found that HCC-derived exosomal miR-761 was taken up by NFs.Moreover,HCC exosomes aff ected the tumor microenvironment by activating NFs via suppressor of cytokine signaling 2(SOCS2)and the JAK2/STAT3 signaling pathway.CONCLUSIONS:These results demonstrated that exosomal miR-761 modulated the tumor microenvironment via SOCS2/JAK2/STAT3 pathway-dependent activation of CAFs.Our fi ndings may inspire new strategies for HCC prevention and therapy.

Inhibition of cerebral ischemia/reperfusion injuryinduced apoptosis:nicotiflorin and JAK2/STAT3 pathway

Nicotiflorin is a flavonoid extracted from Carthamus tinctorius.Previous studies have shown its cerebral protective effect,but the mechanism is undefined.In this study,we aimed to determine whether nicotiflorin protects against cerebral ischemia/reperfusion injury-induced apoptosis through the JAK2/STAT3 pathway.The cerebral ischemia/reperfusion injury model was established by middle cerebral artery occlusion/reperfusion.Nicotiflorin（10 mg/kg） was administered by tail vein injection.Cell apoptosis in the ischemic cerebral cortex was examined by hematoxylin-eosin staining and terminal deoxynucleotidyl transferase d UTP nick end labeling assay.Bcl-2 and Bax expression levels in ischemic cerebral cortex were examined by immunohistochemial staining.Additionally,p-JAK2,p-STAT3,Bcl-2,Bax,and caspase-3 levels in ischemic cerebral cortex were examined by western blot assay.Nicotiflorin altered the shape and structure of injured neurons,decreased the number of apoptotic cells,down-regulates expression of p-JAK2,p-STAT3,caspase-3,and Bax,decreased Bax immunoredactivity,and increased Bcl-2 protein expression and immunoreactivity.These results suggest that nicotiflorin protects against cerebral ischemia/reperfusion injury-induced apoptosis via the JAK2/STAT3 pathway.

KIAA1199 induces advanced biological behavior and development of ovarian cancer through activation of the IL-6/STAT3 pathway

Recently,abnormal expression of KIAA1199 has been detected in Epithelial Ovarian Cancer(EOC).However,the underlined anti-ovarian cancer mechanism of KIAA1199 remains to be enlightened.In our study,we performed to elucidate the effects of KIAA1199 on the advanced biological behavior of EOC cells through activation of the IL-6/STAT3 pathway.Confirmed by immunohistochemistry,KIAA1199 was highly expressed in ovarian borderline and malignant epithelial tumors.A retrospective analysis found that EOC patients with low expression of KIAA1199 had a significantly higher 5-year survival rate than those with high expression.Mechanistically,IL-6 was used to stimulate EOC cells,and the expression of KIAA1199,STAT3 and p-STAT3 increased after IL-6 stimulation.These results could show that KIAA1199 is transcriptionally activated by IL6/STAT3 pathway,thereby accelerating the deterioration of EOC.KIAA1199 could also be used as a poor prognosis factor and potential target in treatment.

Value of spiral CT perfusion parameters for evaluating acute pancreatitis and their correlation with inflammatory factor and JAK2/STAT3 signaling pathway

Objective: To study the value of spiral CT perfusion parameters for evaluating acute pancreatitis and their correlation with inflammatory factor and JAK2/STAT3 signaling pathway. Methods: Patients with acute pancreatitis and patients with pancreatic trauma who underwent surgical resection in Liaocheng Dongchangfu People's Hospital between May 2014 and March 2017 were selected and enrolled in the AP group and the control group of the research respectively;spiral CT perfusion scanning was conducted before surgery to measure the blood flow (BF), blood volume (BV), and mean transit time (MTT), and the serum was collected to determine the contents of inflammatory factors;pancreatitis tissue and normal pancreatic tissue were collected after surgical resection to determine the expression of JAK2/STAT3 signal molecules. Results: pancreatic tissue BF and BV levels of AP group were significantly lower than those of control group while MTT level was not different from that of control group;CRP, PCT, HMGB-1, Ghrelin and sTREM-1 contents in serum as well as JAK2, STAT3, Bcl-2 and Bcl-xL mRNA expression in pancreatic tissue of AP group were significantly higher than those of control group and negatively correlated with BF and BV levels in pancreatic tissue. Conclusion: Spiral CT perfusion parameters BF and BV can reflect the microcirculatory disorder of acute pancreatitis and are associated with the increased secretion of inflammatory factors and the activation of JAK2/STAT3 signaling pathway in the course of disease.

GPR160 regulates the self-renewal and pluripotency of mouse embryonic stem cells via JAK1/STAT3 signaling pathway

G-protein-coupled receptors(GPCRs)are the largest family of transmembrane receptors and regulate various physiological and pathological processes.Despite extensive studies,the roles of GPCRs in mouse embryonic stem cells(mESCs)remain poorly understood.Here,we show that GPR160,a class A member of GPCRs,is dramatically downregulated concurrent with mESC differentiation into embryoid bodies in vitro.Knockdown of Gpr160 leads to downregulation of the expression of pluripotency-associated transcription factors and upregulation of the expression of lineage markers,accompanying with the ar-rest of the mESC cell-cycle in the GO/G1 phase.RNA-seq analysis shows that GPR160 participates in the JAK/STAT signaling pathway crucial formaintaining ESC stemness,and the knockdown of Gpr160 results in the downregulation of STAT3 phosphorylation level,which in turn is partially rescued by colivelin,a STAT3 activator.Consistent with these observations,GPR160 physically interacts with JAK1,and co-operates with leukemia inhibitory factor receptor(LIFR)and gp130 to activate the STAT3 pathway.In summary,our results suggest that GPR160 regulates mESC self-renewal and pluripotency by interacting with the JAK1-LIFR-gp130 complex to mediate the JAK1/STAT3 signaling pathway.

Correlation of expression of STAT3,VEGF and differentiation of Th17 cells in psoriasis vulgaris of guinea pig

Objective:To investigate the role of T help 17 cells(Th17)and STAT3-VEGF pathway in pathogenesis of psoriasis.Methods:A total of 50 cases of psoriasis guinea pigs and 20 normal guinea pigs were selected.The ratio of Th17/IL-17 cell in peripheral blood were detected by flow cytometric analysis;STAT3 and VEGF concentrations were measured hy immunohistochemistry and Western blot.Results:The expression of Th17 in peripheral blood were significantly increased in psoriasis[(1.76±0.88)%]compared with controls[(0.48±0.27)%](P<0.05).Th17 related cytokine STAT3 and VEGF were significantly increased in psoriasis compared with controls(P<0.05),and were positively correlated the expression of Th17.Conclusions:The expressions of Th17,STAT3 and VEGF are elevated in psoriasis,which suggests Th17 cells have a potential role in the pathogenesis of psoriasis by STAT3-VEGF pathway.

Puerarin protects rat brain against ischemia/reperfusion injury by suppressing autophagy via the AMPK-mT OR-ULK1 signaling pathway

Puerarin suppresses autophagy to alleviate cerebral ischemia/reperfusion injury, and accumulating evidence indicates that the AMPKm TOR signaling pathway regulates the activation of the autophagy pathway through the coordinated phosphorylation of ULK1. In this study, we investigated the mechanisms underlying the neuroprotective effect of puerarin and its role in modulating autophagy via the AMPK-m TOR-ULK1 signaling pathway in the rat middle cerebral artery occlusion model of cerebral ischemia/reperfusion injury. Rats were intraperitoneally injected with puerarin, 50 or 100 mg/kg, daily for 7 days. Then, 30 minutes after the final administration, rats were subjected to transient middle cerebral artery occlusion for 90 minutes. Then, after 24 hours of reperfusion, the Longa score and infarct volume were evaluated in each group. Autophagosome formation was observed by transmission electron microscopy. LC3, Beclin-1 p62, AMPK, m TOR and ULK1 protein expression levels were examined by immunofluorescence and western blot assay. Puerarin substantially reduced the Longa score and infarct volume, and it lessened autophagosome formation in the hippocampal CA1 area following cerebral ischemia/reperfusion injury in a dose-dependent manner. Pretreatment with puerarin（50 or 100 mg/kg） reduced Beclin-1 expression and the LC3-II/LC3-I ratio, as well as p-AMPK and p S317-ULK1 levels. In comparison, it increased p62 expression. Furthermore, puerarin at 100 mg/kg dramatically increased the levels of p-m TOR and p S757-ULK1 in the hippocampus on the ischemic side. Our findings suggest that puerarin alleviates autophagy by activating the APMK-m TOR-ULK1 signaling pathway. Thus, puerarin might have therapeutic potential for treating cerebral ischemia/reperfusion injury.