黄芪对大黄诱导大鼠腹泻的治疗作用及其机制  

Therapeutic effect and its mechanism of Astragali Radix on rhubarb-induced diarrhea in rats

作　　者：庄雪峰 律广富[1] 林贺[1] 黄晓巍[1] 周佳 李禹墨 赵嘉睿 林喆[1] 王雨辰 ZHUANG Xuefeng;LYU Guangfu;LIN He;HUANG Xiaowei;ZHOU Jia;LI Yumo;ZHAO Jiarui;LIN Zhe;WANG Yuchen(Department of Chinese Pharmacology,School of Pharmacy,Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学药学院中药药理学实验室,吉林长春130117

出　　处：《吉林大学学报（医学版）》2022年第5期1156-1166,共11页Journal of Jilin University：Medicine Edition

基　　金：吉林省发改委资助项目(2020C032-2);吉林省卫健委卫生与健康技术创新项目(2020J068);长春中医药大学杏林学者计划项目(QNKXJ2-2021ZR04)。

摘　　要：目的:探讨黄芪对于大黄诱导腹泻大鼠肠道炎症、肠道屏障和肠道菌群的治疗作用,并阐明其可能的作用机制。方法:50只大鼠随机分为对照组、模型组、阳性对照组(2.468 g·kg^(-1)参苓白术散)、低剂量(1.35 g·kg^(-1))黄芪组和高剂量(2.70 g·kg^(-1))黄芪组,每组10只。采用大黄灌胃结合饮食不节复制大鼠腹泻模型,检测各组大鼠体质量、粪便含水量和腹泻评分;HE染色观察各组大鼠结肠组织病理形态表现,Western blotting法检测各组大鼠结肠组织中蛋白酶激活受体2 (PAR-2)、磷酸化p38 (p-p38)、肌球蛋白轻链激酶(MLCK)、磷酸化肌球蛋白轻链(p-MLC)、闭锁蛋白1(ZO-1)、闭合蛋白(Occludin)、Toll样受体4 (TLR4)、肿瘤坏死因子受体相关因子6 (TRAF6)、髓样分化因子88 (MyD88)和核因子κB (NF-κB)蛋白表达水平;采用酶联免疫吸附测定(ELISA)法检测各组大鼠血清中胃动素(MTL)、胃泌素(GAS)、肿瘤坏死因子α (TNF-α)、白细胞介素1β(IL-1β)、白细胞介素6 (IL-6)、白细胞介素10 (IL^(-1)0)、丙二醛(MDA)和分泌型免疫球蛋白A(SIgA)水平及超氧化物歧化酶(SOD)活性;16S r DNA基因测序分析各组大鼠肠道菌群情况。结结果果:与对照组比较,模型组大鼠体质量明显降低(P<0.01),粪便含水量和腹泻评分明显升高(P<0.01),血清中MTL和GAS水平明显升高(P<0.01);与模型组比较,阳性对照组、低剂量黄芪组和高剂量黄芪组大鼠体质量明显升高(P<0.01),粪便含水量和腹泻评分明显降低(P<0.01),血清中MTL和GAS水平明显降低(P<0.01)。HE染色,与对照组比较,模型组大鼠结肠组织上皮黏膜损伤,有大量炎症细胞浸润,腺体排列紊乱;与模型组比较,阳性对照组和高剂量黄芪组大鼠结肠组织上皮黏膜结构较规则,炎症细胞浸润较少,未见明显腺体紊乱,低剂量黄芪组大鼠结肠组织上皮黏膜结构可见轻微损伤,有部分炎症细胞浸润,腺体排列较整齐。Western blottingObjective:To investigate the therapeutic effects of Astragali Radix on the intestinal inflammation,intestinal barrier and intestinal flora in the rats with rhubarb-induced diarrhea,and to elucidate the possible mechanisms. Methods:Fifty rats were randomly divided into control group,model group,positive control group(2. 468 g·kg^(-1)Shenlingbaizhu Powder),low dose(1. 35 g·kg^(-1))of Astragali Radix group,and high dose(2. 70 g·kg^(-1))of Astragali Radix group,with 10 rats in each group. The rat diarrhea models were replicated by rhubarb gavage in combination with diet indiscipline. The body weights,fecal water contents and diarrhea scores of the rats in various groups were detected;HE staining was used to observe the pathomorphology of colon tissue of the rats in various groups,and Western blotting method was used to detect the expression levels of protease activated receptor-2(PAR-2),phosphorylated p38(p-p38),myosin light chain kinase(MLCK),phosphorylated myosin light chain(p-MLC),zonula occludens-1(ZO-1),Occludin,Toll-like receptor 4(TLR4),tumor necrosis factor receptor-associated factor 6(TRAF6),myeloid differentiation factor 88(MyD88),and nuclear factor-κB(NF-κB)proteins in colon tissue of the rats in various groups;the levels of motilin(MTL),gastrin(GAS),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β), interleukin-6(IL-6),interleukin-10(IL^(-1)0),malondialdehyde(MDA),and secretory immunoglobulin A(SIgA)and the activities of superoxide dismutase(SOD)in serum of the rats in various groups were measured by enzyme linked immunosorbent assay(ELISA)method;16S r DNA gene sequencing was performed to analyze the intestinal flora of the rats in various groups. Results:Compared with control group,the body weight of the rats in model group was significantly decreased(P<0. 01),the fecal water content and diarrhea score were significantly increased(P<0. 01),and the levels of MTL and GAS in serum were significantly increased(P<0. 01);compared with model group,the body weights of the rats in positive control and

关 键 词：黄芪 肠道屏障 肠道炎症 肠道菌群 革兰阴性菌 

分 类 号：R285.5[医药卫生—中药学]