摘要
目的:研究安肠汤低、中、高剂量对SD大鼠溃疡性结肠炎的抗炎作用,通过研究不同给药剂量对miRNA-146a/非受体型酪氨酸蛋白激酶(JAK)/信号传导与转录激活因子(STAT)/细胞因子信号传导抑制蛋白-3(SOCS-3)信号通路及其下游蛋白的影响探讨安肠汤防治溃疡性结肠炎的可能机制。方法:实验大鼠分为正常组,模型组,美沙拉嗪组(1 g·kg^(-1)),安肠汤低、中、高剂量组(6,12,24 g·kg^(-1)),每组10只。除正常组外,其余各组均采用2,4,6-三硝基苯磺酸(TNBS)/乙醇灌肠法建立溃疡性结肠炎大鼠模型,分别给药14 d。观察大鼠神态、大便性状、毛发等一般情况的变化,并参照疾病活动指数(DAI)表进行评分,苏木素-伊红(HE)染色观察各组大鼠结肠组织病理改变,酶联免疫吸附测定(ELISA)检测大鼠血清肿瘤坏死因子-α(TNF-α),白细胞介素-10(IL-10),IL-17,IL-1β,IL-6水平,蛋白免疫印迹法(Western blot)检测结肠组织中JAK2,磷酸化STAT3(p-STAT3),STAT3,SOCS-3蛋白表达的变化,通过实时荧光定量聚合酶链式反应(Real-time PCR)测定大鼠结肠组织中JAK2,p-STAT3,STAT3,SOCS-3 mRNA以及血浆miRNA-146a的表达水平。结果:与正常组比较,模型组大鼠JAK2,p-STAT3,STAT3蛋白表达及JAK2,p-STAT3,STAT3 mRNA表达明显增高(P<0.05),模型组大鼠miRNA-146a,SOCS-3 mRNA以及SOCS-3蛋白表达明显降低(P<0.05);与模型组比较,给药组大鼠精神状态,进食量,毛色等情况均明显改善,DAI评分明显降低(P<0.05),给药组大鼠结肠溃疡组织明显改善,给药组大鼠结肠组织JAK2,p-STAT3,STAT3蛋白表达及JAK2,p-STAT3,STAT3 mRNA表达明显降低(P<0.05),给药组大鼠miRNA-146a,SOCS-3 mRNA以及SOCS-3蛋白表达明显增高(P<0.05)。结论:安肠汤可能通过影响miRNA-146a/JAK/STAT/SOCS-3信号转导通路相关基因及蛋白表达,抑制溃疡性结肠炎病情的发展。
Objective: To study the anti-inflammatory effects of low,middle,and high doses of Anchang decoction on ulcerative colitis in SD rats,and also explore the possible mechanism of Anchang decoction in the prevention and treatment of ulcerative colitis through the effect of different doses on miRNA-146 a/non-receptor tyrosine protein kinase(JAK)/signal transduction and activator of transcription 3(STAT3)/cytokine signaling protein-3(SOCS-3)signal pathway and its downstream proteins. Method: The experimental rats were divided into control group,model group,mesalazine group(1 g·kg^(-1))and Anchang decoction low(6 g·kg^(-1)),middle(12 g·kg^(-1))and high dose groups(24 g·kg^(-1)),with 10 rats in each group. Except for the control group,2,4,6-trinitrobenzenesulfonic acid(TNBS)/ethanol enema was used in all the other groups to establish a rat model of ulcerative colitis for 14 days respectively. The general changes of the mental state,stool traits,hair and other general conditions of the rats were observed,and score was graded with reference to the disease activity index(DAI)table. The pathological changes of colon tissue of rats in each group were observed by hematoxylin-eosin(HE)staining. The levels of serum tumor necrosis factor-α(TNF-α),interleukin-10(IL-10),interleukin-17(IL-17),interleukin-1β(IL-1β)and interleukin-6(IL-6)were detected by enzyme linked immunosorbent assay(ELISA). The expression levels of JAK2,phosphorylation STAT3(p-STAT3),STAT3 and inhibitor of SOCS-3 in colon tissue were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the expression of JAK2,p-STAT3,STAT3,SOCS-3 mRNA in rat colon and miRNA-146 a in rat plasma. Result: Compared with the normal group,the expression of JAK2,p-STAT3,STAT3 protein and the expression of JAK2,p-STAT3 and STAT3 mRNA in the model group increased(P<0.05),and the relative expression of miRNA-146 a,SOCS-3 mRNA and SOCS-3 protein decreased in the model group(P<0.05). Compared with the model group,the
作者
梁运特
孙平良
LIANG Yun-te;SUN Ping-liang(Postgraduate College of Guangxi Chinese Medicine University,Nanning 530200,China;The First Affiliated Hospital of Guangxi Chinese Medicine University,Nanning 530023,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第3期30-38,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81660795)。
