黄芪总苷通过PAR2信号通路减轻糖尿病大鼠肝纤维化的机制研究  

Study on the mechanism of total glycosides of astragalus in reducing liver fibrosis in diabetic rats through PAR2 signaling pathway

作　　者：李芬 王甲彬 张世龙 左麦红 LI Fen;WANG Jia-bin;ZHANG Shi-long(Department of Hepatobiliary Surgery,Qingyang Gansu 745000,China;Department of Endocrinology,Second People's Hospital of Qingyang City,Qingyang Gansu 745000,China;School of Public Health,Lanzhou University,Lanzhou Gansu 730000,China.)

机构地区：[1]庆阳市第二人民医院肝胆外科,甘肃庆阳745000 [2]庆阳市第二人民医院内分泌科,甘肃庆阳745000 [3]庆阳市第二人民医院检验科,甘肃庆阳745000 [4]兰州大学公共卫生学院,甘肃兰州730000

出　　处：《临床和实验医学杂志》2021年第10期1043-1047,共5页Journal of Clinical and Experimental Medicine

基　　金：甘肃省青年基金项目(编号:150RJYM325)。

摘　　要：目的探讨黄芪总苷在蛋白酶激活受体2(PAR2)信号通路的作用下缓解糖尿病大鼠肝纤维化的作用机制。方法采取随机数字表法分组,将100只大鼠平均分为5组,各20只,分别为空白对照组、模型组、黄芪总苷低、中、高剂量组。除空白对照组,其他各组均采用一次性腹腔注射构建糖尿病大鼠模型。空白对照组和模型组大鼠给予0.5 mg/kg 0.9%氯化钠溶液灌胃,黄芪总苷低、中、高剂量组大鼠分别给予0.5、1.0、2.0 mg/kg黄芪总苷灌胃。各组大鼠均连续灌胃28 d后,采用自动生化分析仪检测各组大鼠天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、血清白蛋白(ALB)和白球蛋白比例(A/G)等肝脏功能指标;采用蛋白质印迹法检测各组大鼠肝组织蛋白Smad 2、Smad 3、pSmad 2、pSmad 3和α-SMA以及PAR2信号通路蛋白PAR2、P-P38 MAPK和P38 MAPK的表达水平;采用放射免疫法测定各大鼠肝脏组织透明质酸(HA)、层黏连蛋白(LN)、Ⅳ型胶原(Ⅳ-C)和Ⅲ型前胶原(PC-Ⅲ)的水平;采用酶联免疫吸附试验(ELISA)检测大鼠血清白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和转化生长因子β1(TGF-β1)水平。结果与空白对照组相比,模型组大鼠AST和ALT、肝组织蛋白、HA和LN、炎性细胞因子、TGF-β1和PAR2信号通路蛋白水平均显著升高,ALB、A/G、Ⅳ-C和PC-Ⅲ水平则显著降低,差异均有统计学意义(P<0.05);黄芪总苷灌胃治疗后,高剂量组作用效果最为显著,表现为:与模型组相比,大鼠AST和ALT、肝组织蛋白、HA和LN、炎性细胞因子、TGF-β1和PAR2信号通路蛋白水平均显著降低,ALB、A/G、Ⅳ-C和PC-Ⅲ水平则显升高,差异均有统计学意义(P<0.05)。结论黄芪总苷可通过调节糖尿病大鼠肝脏功能指标和肝组织蛋白水平,并可改善纤维化程度,降低炎症反应水平,从而减轻糖尿病大鼠肝纤维化并改善病情,其发挥作用的相关机制与PAR2信�Objective To investigate the mechanism of astragaloside alleviating liver fibrosis in diabetic rats under the action of PAR2 signaling pathway.Methods A random number table method was adopted to group,and 100 rats were divided into 5 groups,each with 20 rats,which were the blank control group,the model group,and the low and medium-high-dose groups of total astragalus glycosides.Except for the blank control group,all other groups used one-time intraperitoneal injection to construct a diabetic rat model.The rats in the blank control group and the model group were given 0.5 mg/kg normal saline by gavage,and the rats in the low,middle and high-dose groups of astragaloside were given 0.5,1.0,2.0 mg/kg total astragaloside respectively.After continuous intragastric administration for 28 days,the liver functions indicators such as aspartate aminotransferase(AST),alanine aminotransferase(ALT),serum albumin(ALB)and albumin ratio(A/G)in each group of rats were tested using automatic biochemical analyzer;the expression levels of rat liver tissue proteins Smad 2,Smad 3,pSmad 2,pSmad 3,α-SMA,and PAR2 signaling pathway proteins PAR2,P-P38 MAPK,and P38 MAPK in each group were detected by Western blotting;the levels of hyaluronic acid(HA),laminin(LN),collagen typeⅣ(Ⅳ-C)and procollagen typeⅢ(PC-Ⅲ)in the liver tissue were determined by radioimmunoassay;the levels of serum interleukin-6(interleukin-6,IL-6),interleukin-1β(interleukin-1β,IL-1β),tumor necrosis factorα(TNF-α)and transforming growth factorβ1(TGF-β1)were detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with the control group,the levels of AST and ALT,liver tissue protein expression,HA and LN,inflammatory cytokine,pro-fibrosis factor and PAR2 signaling pathway protein in the model group were significantly increased,while the levels of ALB,A/G,Ⅳand PCⅢwere significantly reduced,the differences were statistically significant(P<0.05);After gavage treatment with total astragalus glycosides,the high-dose group has the most significant ef

关 键 词：大鼠 黄芪总苷 PAR2信号通路 糖尿病 肝纤维化 促纤维化因子 

分 类 号：R285.5[医药卫生—中药学]