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.

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.

Hypoglycemic mechanism of Tegillarca granosa polysaccharides on type 2 diabetic mice by altering gut microbiota and regulating the PI3K-akt signaling pathwaye

Type 2 diabetes mellitus(T2DM)is a complex metabolic disease threatening human health.We investigated the effects of Tegillarca granosa polysaccharide(TGP)and determined its potential mechanisms in a mouse model of T2DM established through a high-fat diet and streptozotocin.TGP(5.1×10^(3) Da)was composed of mannose,glucosamine,rhamnose,glucuronic acid,galactosamine,glucose,galactose,xylose,and fucose.It could significantly alleviate weight loss,reduce fasting blood glucose levels,reverse dyslipidemia,reduce liver damage from oxidative stress,and improve insulin sensitivity.RT-PCR and Western blotting indicated that TGP could activate the phosphatidylinositol-3-kinase/protein kinase B signaling pathway to regulate disorders in glucolipid metabolism and improve insulin resistance.TGP increased the abundance of Allobaculum,Akkermansia,and Bifidobacterium,restored the microbiota abundance in the intestinal tracts of mice with T2DM,and promoted short-chain fatty acid production.This study provides new insights into the antidiabetic effects of TGP and highlights its potential as a natural hypoglycemic nutraceutical.

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.

Endogenous hydrogen sulfide and ERK1/2-STAT3 signaling pathway may participate in the association between homocysteine and hypertension

Background Homocysteine(Hcy)is a risk factor for hypertension,although the mechanisms are poorly understood.Methods We first explored the relationship between Hcy levels and blood pressure(BP)by analyzing the clinical data of primary hypertensive patients admitted to our hospital.Secondly,we explored a rat model to study the effect of Hcy on blood pressure and the role of H2S.An hyperhomocysteinemia(HHcy)rat model was induced to explore the effect of Hcy on blood pressure and the possible mechanism.We carried out tissue histology,extraction and examination of RNA and protein.Finally,we conducted cell experiments to determine a likely mechanism through renin-angiotensin-aldosterone system(RAAS)and extracellular signal-regulated kinase 1/2(ERK1/2)signaling pathway.Results In primary hypertensive inpatients with HHcy,blood pressure was significantly higher as compared with inpatient counterparts lacking HHcy.In the rat model,blood pressure of the Wistar rats was significantly increased with increases in serum Hcy levels and decreased after folate treatment.Angiotensin converting enzyme 1(ACE1)expression in the Wistar Hcy group was enhanced comparing to controls,but was decreased in the Wistar folate group.Angiotensin II receptor type 1(AGTR1)levels in the kidney tissue increased in the Wistar folate group.Both serum H2S and kidney cystathionineγ-lyase decreased with elevated levels of serum Hcy.In vitro,increased concentrations and treatment times for Hcy were associated with increased expression of collagen type 1 and AGTR1.This dose and time dependent response was also observed for p-STAT3 and p-ERK1/2 expression.Conclusion Endogenous H2S might mediate the process of altered blood pressure in response to changes in serum Hcy levels,in a process that is partly dependent on activated RAAS and ERK1/2-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.

基于JAK2-STAT3信号通路探讨热毒宁注射液治疗大鼠急性肺损伤的作用机制

目的:探讨热毒宁注射液通过调节JAK2-STAT3信号通路干预急性肺损伤(ALI)大鼠作用机制。方法:30只Wistar大鼠随机分为五组,分别为模型对照组、空白对照组、地塞米松组、热毒宁高剂量组和热毒宁低剂量组。每组大鼠均进行腹腔注射给药,每日1次,连续5 d,末次给药后,采用脂多糖滴注咽后壁气管建立ALI大鼠模型。造模24 h后,比较各组大鼠动脉氧分压,HE染色法观察肺组织的病理学改变,ELISA法检测大鼠肺组织肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的表达水平,Western blot测定各组大鼠肺组织JAK2、p-JAK2、STAT3、p-STAT3蛋白表达水平。结果:与空白对照组比较,模型对照组大鼠PaO 2水平明显下降,病理评分升高,PaCO 2、IL-1β、IL-6、TNF-α水平、JAK2、p-JAK2、STAT3、p-STAT3蛋白表达升高(均P<0.05)。与模型对照组比较,地塞米松组和热毒宁高剂量组大鼠PaO 2水平升高,病理评分下降,PaCO 2、IL-1β、IL-6、TNF-α水平、JAK2、p-JAK2、STAT3、p-STAT3蛋白表达降低(均P<0.05)。结论:热毒宁注射液对ALI大鼠有保护作用,其机制可能与调节JAK2-STAT3信号通路的活化、降低炎症反应有关。

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.

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.

MiR-19a-3p regulates the Forkhead box F2-mediated Wnt/β-catenin signaling pathway and affects the biological functions of colorectal cancer cells

BACKGROUND Colorectal cancer(CRC)is one of the most common malignancies worldwide.AIM To explore the expression of microRNA miR-19a-3p and Forkhead box F2(FOXF2)in patients with CRC and the relevant mechanisms.METHODS Sixty-two CRC patients admitted to the hospital were enrolled into the study group,and sixty healthy people from the same period were assigned to the control group.Elbow venous blood was sampled from the patients and healthy individuals,and blood serum was saved for later analysis.MiR-19a-3p mimics,miR-19a-3p inhibitor,miR-negative control,small interfering-FOXF2,and short hairpin-FOXF2 were transfected into HT29 and HCT116 cells.Then quantitative polymerase chain reaction was performed to quantify the expression of miR-19a-3p and FOXF2 in HT29 and HCT116 cells,and western blot(WB)analysis was conducted to evaluate the levels of FOXF2,glycogen synthase kinase 3 beta(GSK-3β),phosphorylated GSK-3β(p-GSK-3β),β-catenin,p-β-catenin,α-catenin,Ncadherin,E-cadherin,and vimentin.The MTT,Transwell,and wound healing assays were applied to analyze cell proliferation,invasion,and migration,respectively,and the dual luciferase reporter assay was used to determine the correlation of miR-19a-3p with FOXF2.RESULTS The patients showed high serum levels of miR-19a-3p and low levels of FOXF2,and the area under the curves of miR-19a-3p and FOXF2 were larger than 0.8.MiR-19a-3p and FOXF2 were related to sex,tumor size,age,tumor-nodemetastasis staging,lymph node metastasis,and differentiation of CRC patients.Silencing of miR-19a-3p and overexpression of FOXF2 suppressed the epithelialmesenchymal transition,invasion,migration,and proliferation of cells.WB analysis revealed that silencing of miR-19a-3p and FOXF2 overexpression significantly suppressed the expression of p-GSK-3β,β-catenin,N-cadherin,and vimentin;and increased the levels of GSK-3β,p-β-catenin,α-catenin,and Ecadherin.The dual luciferase reporter assay confirmed that there was a targeted correlation of miR-19a-3p with FOXF2.In addition,a rescue experiment revealed that there were no differences in cell proliferation,invasion,and migration in HT29 and HCT116 cells co-transfected with miR-19a-3p-mimics+sh-FOXF2 and miR-19a-3p-inhibitor+si-FOXF2 compared to the miR-negative control group.CONCLUSION Inhibiting miR-19a-3p expression can upregulate the FOXF2-mediated Wnt/β-catenin signaling pathway,thereby affecting the epithelial-mesenchymal transition,proliferation,invasion,and migration of cells.Thus,miR-19a-3p is likely to be a therapeutic target in CRC.

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.

β-榄香烯阻断JAK2-STAT3信号通路促进紫杉醇对肺癌细胞增殖和凋亡作用研究

目的:研究分析讨论β-榄香烯对紫杉醇抑制肺癌细胞增殖、诱导凋亡的作用,并探讨其可能的机制。方法:建立耐药细胞株A549/Taxol,MTT法测定β-榄香烯对A549/Taxol细胞株的抑制率及IC50;使用Annexin V-FITC/PI检测细胞凋亡率;并通过Western Blot法检测JAK2、STAT3、p-STAT3、Bcl-2、Bax、Caspase3蛋白的表达水平,借以分析β-榄香烯对紫杉醇抑制肺癌细胞增殖、诱导凋亡的作用和机制。结果:药物干预48 h后,可见β-榄香烯对A549/Taxol肺癌细胞活性均显示出抑制作用,并且抑制作用表现出明显的剂量依赖性,对肺癌细胞抑制的IC50水平为108.5μg/mL。研究中采用IC50浓度108.5μg/mL榄香烯干预A549/Taxol肺癌细胞, 24 h后可见A549/Taxol肺癌细胞的凋亡水平显著增加(P<0.05);与此同时,肺癌细胞JAK2、STAT3、p-STAT3和Bcl-2可见降低(P<0.05),而Bax和Caspase3则见升高(P<0.05)。结论:β-榄香烯可能通过抑制JAK2/STAT3信号通路,发挥拮抗肺癌细胞对紫杉醇的耐药性作用,抑制肿瘤细胞增殖,诱导凋亡。

IL-1β通过JAK2-STAT3促进大鼠脊髓损伤后胶质瘢痕形成

目的探讨IL-1β促进脊髓损伤后胶质瘢痕形成的机制。方法将大鼠随机分为模型组(采用钳夹脊髓的方法建立SCI模型)、假手术组(sham group)、IL-1β特异性抑制剂组(IL-1RA)、IL-1β组(IL-1β)及IL-1β+JAK2-STAT3特异性抑制剂组(IL-1β+AG490)。假手术组只打开椎板,不作其他处理。在术后相应时间点(术后8及12 h和1、3、7及14 d)进行大鼠后肢BBB评分,用Western blot、免疫荧光和免疫组化技术检测GFAP、vimentin、p-STAT3的表达变化。结果 p-STAT3(术后第8 h和第12 h)及GFAP、vimentin(术后第7和第14天)表达趋势:模型组显著高于假手术组(P<0.01),IL-1RA组明显低于模型组(P<0.05),但仍高于假手术组(P<0.05);IL-1β+AG490组明显低于模型组(P<0.05),但仍高于假手术组(P<0.05);IL-1β组均显著高于模型组(P<0.05)。术后第14天,BBB评分模型组显著低于假手术组(P<0.01),IL-1RA组显著高于模型组(P<0.05),但仍低于假手术组(P<0.01);IL-1β组显著低于模型组(P<0.05)。结论 IL-1β可通过JAK2-STAT3促进脊髓损伤后胶质瘢痕形成,抑制IL-1β或JAK2-STAT3可减弱胶质瘢痕形成,促进脊髓神经功能恢复。

保肝宁对瘦素刺激HSC增殖及其JAK_2-STAT_3信号通路影响的实验研究

目的观察中药复方保肝宁对瘦素刺激肝星状细胞(hepatic stellate cell,HSC)增殖功能的作用及其对JAK2-STAT3信号通路的影响。方法给正常Wistar大鼠灌胃保肝宁煎剂7d,制备含药血清,用噻唑兰(MTT)比色法检测保肝宁对100ng/ml的瘦素刺激HSC-LX2增殖的影响;应用蛋白印迹试验检测保肝宁对100ng/ml的瘦素刺激HSC-LX2中的JAK2、STAT3蛋白的表达水平。结果保肝宁组对瘦素刺激的HSC-LX2增殖有显著的抑制作用;保肝宁作用6h后,JAK2、STAT3蛋白表达水平开始降低,此时JAK2降低较为明显;24h时STAT3蛋白降低较为明显。结论中药复方保肝宁有抑制瘦素促HSC-LX2增殖的作用,而JAK2-STAT3通路被阻断,JAK2、STAT3蛋白表达的降低可能是其发生的主要机制之一,而这一作用主要是通过阻断瘦素发生生物学效应的JAK2-STAT3经典通路,降低JAK2、STAT3蛋白表达来实现的。

JAK2-STAT3信号通路在舒芬太尼预处理诱导大鼠心肌保护效应中的作用

目的 探讨舒芬太尼预处理对大鼠心肌缺血再灌注损伤的影响以及JAK2-STAT3信号通路的作用.方法 雄性SD大鼠60只,体质量250～300 g,随机均分为假手术组（S组）、缺血再灌注组（I/R组）、舒芬太尼预处理组（SPC组）、舒芬太尼预处理联合JAK2激酶抑制剂组（S＋A组）及JAK2激酶抑制剂组（A组）.采用结扎冠状动脉左前降支的方法制作心肌缺血再灌注模型.SPC组于心肌缺血前股静脉泵注舒芬太尼1μg/kg,泵注5 min,间隔5 min,重复处理3次,总量共3 μg/kg;S＋A组：舒芬太尼预处理前5 min给予JAK2激酶抑制剂AG490（1 mg/kg）,缺血前30min舒芬太尼预处理;A组：缺血前35 min给予JAK2激酶抑制剂AG490（1 mg/kg）.心肌缺血前30min（T0）、缺血前即刻（T1）、缺血30 min（T2）、再灌注30 min（T3）和再灌注120 min（T4）时记录心率（HR）和平均动脉压（MAP）.再灌注120 min抽取动脉血,离心取血清测定肌酸激酶同工酶（CK-MB）和乳酸脱氢酶（LDH）.实验结束时,各组取6只大鼠心脏测算心肌梗死面积,其余6只大鼠采用Western blot测定心肌组织磷酸化STAT3 （P-STAT3）的表达.结果 比较各组间不同时点HR的差异无统计学意义（P＞0.05）.与S组相比,其余各组T2～T4时MAP降低（P＜0.05或P＜0.01）;血清CK-MB和LDH活性明显增高（P＜o.01）.与I/R组相比,SPC组MAP数值稍高,但差异无统计学意义;CK-MB和LDH活性降低（P＜0.01）;心肌梗死范围减小（P＜0.01）.与S组比较,I/R组和SPC组P-STAT3表达上调（P＜0.01）,且SPC组上调高于I/R组（P＜0.01）.结论 舒芬太尼预处理减轻大鼠心肌缺血再灌注损伤的机制可能与激活JAK2-STAT3信号通路、上调P-STAT3的表达有关.

JAK_2-STAT_3信号通路在白介素-1β经动脉外膜给药致平滑肌细胞增殖迁移中的作用

目的研究白细胞介素-1β经血管外膜给药导致动脉平滑肌细胞发生的改变及其与JAK2-STAT3信号通路的关系,明确JAK2-STAT3信号通路在血管增殖性病变中的作用。方法 6～8周龄SD雄性大鼠24只,分离左侧颈总动脉,实验组(n=18)血管外膜包裹含白介素-1β2.5μg的琼脂缓释悬液,对照组(n=6)包裹不含白介素的琼脂悬液,术后2、8、24、48 h,1、2周分别处死实验组大鼠3只,对照组1只,血管标本经切片HE染色观察形态,Western blot法检测JAK2、STAT3、磷酸化JAK2以及磷酸化STAT3的表达,免疫组化标记定位磷酸化JAK2及磷酸化STAT3;另取SD雄性大鼠9只,分离两侧颈总动脉,左侧滴加JAK2抑制剂AG490缓释凝胶,右侧滴加等量空白凝胶后两侧均包裹等量白介素-1β,术后8、48 h,1周处死动物分别行HE染色及Western blot检测。结果白介素-1β包裹血管外膜后出现血管平滑肌细胞的增殖、迁移,通道蛋白JAK2、STAT3在不同时间点出现不同程度的磷酸化,经Western blot检测并行灰度分析,p-JAK2相对灰度值对照组(0.337±0.216),8 h组(1.764±0.513),1周组(0.451±0.229);p-STAT3相对灰度值对照组(0.125±0.870),24 h组(1.909±0.309),2周组(0.448±0.516),各组间有明显差异(P<0.05)。加用抑制剂后,8 h组p-JAK2相对灰度值实验侧(0.085±0.031),对照侧(1.416±0.468),48 h组p-STAT3相对灰度值实验侧(0.460±0.065),对照侧(2.425±0.638),提示JAK2、STAT3的磷酸化水平被明显抑制(P<0.05),平滑肌细胞变化程度下降。结论炎性因子白细胞介素-1β经动脉外膜给药可致动脉平滑肌细胞增殖、迁移,这种改变与JAK2-STAT3信号通路有直接关系。

JAK2-STAT3通路介导肢体缺血后处理脑缺血再灌注损伤保护作用的研究

目的探讨肢体缺血后处理对大鼠脑缺血再灌注损伤后脑保护的作用机制。方法健康雄性SD大鼠48只,随机分为4组即假手术组(sham)、缺血再灌注组(I/R组)、肢体缺血后处理组(Lpost组)、肢体缺血后处理+AG490组(AG组)。I/R组、Lpost组、AG组均做缺血2 h再灌注24 h,Lpost组:再灌注前实施肢体缺血后处理(缺血15 min,灌注15 min)3个循环,AG组:再灌注前5 min时腹腔注射AG490(l mg/kg),其他处理与Lpost组相同。各组大鼠神经功能评分后,采用TTC染色测脑梗死体积,TUNEL法测定脑细胞凋亡。结果 1与sham组相比,I/R组大鼠神经功能缺损加重(P<0.05);与I/R组相比,Lpost组大鼠神经功能缺损减轻(P<0.05);与Lpost组相比,AG组大鼠神经功能缺损加重(P<0.05);2 sham组大鼠无梗死灶,与I/R组相比,Lpost组大鼠梗死体积减小(P<0.05);与Lpost组相比,AG组梗死体积增大(P<0.05);3与I/R组相比,Lpost组大鼠脑细胞凋亡率明显降低(P<0.05);与Lpost组相比,AG组脑细胞凋亡率明显增高(P<0.05)。结论肢体缺血后处理具有显著的脑保护作用,这一作用是由JAK2-STAT3通路介导的。

阿曼托双黄酮通过JAK2-STAT3通路影响甲状腺癌SW579细胞的增殖和凋亡

目的:探讨阿曼托双黄酮(AF)对甲状腺癌SW579细胞中JAK2-STAT3通路活化及其细胞增殖和凋亡的影响。方法:用0、50、100、150、200μmol/L的AF处理SW579细胞24、48、72 h,采用CCK-8和Celigo计数、FCM、WB及qPCR法检测AF对SW579细胞的增殖、凋亡、JAK2-STAT3通路活化及其下游调控基因c-Myc、Bcl2、survivin的mRNA及蛋白表达水平的影响。结果:AF处理后,SW579细胞增殖能力显著下降(P<0.05)且呈浓度依赖性,细胞凋亡呈浓度依赖性增多(P<0.05),细胞中JAK2-STAT3通路的活化受到显著抑制(P<0.05),其下游基因c-Myc、Bcl2、survivin的mRNA及蛋白表达均明显下降(均P<0.05)。结论:AF可通过抑制SW579细胞中JAK2-STAT3通路活化及其下游基因的表达而抑制SW579细胞的增殖并促进其凋亡,有望成为治疗甲状腺癌的有效药物。

加味四逆散对大鼠肝星状细胞株HSC-T6JAK2-STAT3信号通路的影响

目的观察加味四逆散对大鼠肝星状细胞株HSC-T6 JAK2-STAT3信号通路的影响。方法正常大鼠灌胃加味四逆散药物煎剂7 d,制备含药血清;应用蛋白印迹试验检测加味四逆散对100 mg/L瘦素刺激HSC-T6中JAK2、STAT3蛋白的表达水平。结果加味四逆散作用6 h后,JAK2、STAT3蛋白表达水平开始降低,且JAK2降低较为明显,24 h STAT3降低最明显。结论加味四逆散能降低增殖后的肝星状细胞JAK2、STAT3蛋白表达,并阻断JAK2-STAT3经典通路的信号转导作用,这可能是加味四逆散抗肝纤维化的作用之一。