肝宝胶囊对非酒精性脂肪性肝病模型大鼠肠黏膜屏障和肠道菌群的影响
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篇名: 肝宝胶囊对非酒精性脂肪性肝病模型大鼠肠黏膜屏障和肠道菌群的影响
TITLE: Effects of Ganbao capsules on intestinal mucosal barrier and gut microbiota in rats with non-alcoholic fatty liver disease
摘要: 目的 研究肝宝胶囊对非酒精性脂肪性肝病(NAFLD)模型大鼠肠黏膜屏障和肠道菌群的影响,探讨其防治NAFLD的机制。方法将26只SD大鼠随机选择8只作为空白组,给予普通饲料喂养,剩余18只喂食高脂饲料建立NAFLD模型(2只用于成模检查),造模成功后,按随机数字表法分为模型组和肝宝组,每组8只。肝宝组大鼠灌胃肝宝胶囊溶液(1440mg/kg),空白组与模型组大鼠灌胃等体积蒸馏水,每日1次,连续5周。采用全自动生化分析仪检测大鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和甘油三酯(TG)含量;酶联免疫吸附测定法检测大鼠血清中脂多糖、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-1β含量;苏木精-伊红染色法观察大鼠肝脏和回肠组织病理形态;免疫组化法测定回肠组织中闭合蛋白(Occludin)和闭锁小带蛋白-1(ZO-1)的表达;16S核糖体RNA基因测序技术测定大鼠肠道菌群。结果与模型组比较,肝宝组大鼠血清中ALT、AST、TG、脂多糖、TNF-α、IL-6与IL-1β含量均显著降低(P<0.01),肝脏和回肠组织病变程度均明显改善,回肠组织中Occludin和ZO-1蛋白表达水平均显著升高(P<0.01)。肠道菌群分析显示,与模型组比较,肝宝胶囊能提高大鼠肠道菌群的丰富度和多样性;在门水平上,肝宝胶囊能显著升高拟杆菌门相对丰度,降低厚壁菌门相对丰度以及二者相对丰度比值(P<0.01);在属水平上,肝宝胶囊能显著升高乳杆菌属、布劳特氏菌属、拟杆菌属和阿克曼菌属的相对丰度,显著降低普雷沃氏菌属、苏黎世杆菌属、魏斯氏菌属、SMB53和脱硫弧菌属的相对丰度(P<0.05或P<0.01)。各组大鼠肠道菌群间均有差异物种。结论肝宝胶囊可能通过保护肠黏膜屏障功能和调节肠道益生菌/有害菌结构来改善NAFLD。
ABSTRACT: OBJECTIVE To study the effects of Ganbao capsules on intestinal mucosal barrier and gut microbiota in rats with non-alcoholic fatty liver disease (NAFLD), and to explore its mechanism of prevention and treatment of NAFLD. METHODS Eight of 26 SD rats were randomly selected as blank group and fed with ordinary diet, and the remaining 18 rats were fed with high diet to establish NAFLD model (2 for modeling inspection); after successful modeling, they were divided into model group and Ganbao group, with 8 rats in each group. Ganbao group were given Ganbao capsules solution (1 440 mg/kg) intragastrically, and the blank group and model group were given the constant volume of distilled water intragastrically, once a day, for consecutive 5 weeks. The contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and triglyceride (TG) in serum of rats were detected by automatic analyzer; the contents of lipopolysaccharide, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β in serum of rats were detected by enzyme-linked immunosorbent assay. The pathological morphology of liver and ileum tissues were observed by HE staining, the expressions of Occludin and zonula occludens-1 (ZO-1) were detected by immunohistochemistry method, and the intestinal flora were detected by 16S ribosomal RNA gene sequencing technology. RESULTS Compared with the model group, the serum contents of ALT, AST, TG, lipopolysaccharide, TNF-α, IL-6 and IL-1β in Ganbao group were decreased significantly (P<0.01), the pathological changes of liver and ileum tissues were improved 262 significantly, and the expressions of Occludin and ZO-1 were increased significantly (P<0.01). Intestinal microbiotaanalysis revealed that compared with the model group, Ganbao capsules could recover the abundance and diversity of the gut E-mail:hdf8833@126.com microbiota in rats. At the phylum level, Ganbao capsules could significantly increase the relative abundance of Bacteroidetes, and significantly reduce the relative abundance of Firmicutes and the ratio of Firmicutes to Bacteroidetes (P<0.01). At the genus level, Ganbao capsules could significantly increase the relative abundance of Lactobacillus, Blautia, Bacteroides and Akkermansia, and significantly reduce the relative abundance of Prevotella, Turicibacter, Weissella, SMB53 and Desulfovibrio (P<0.05 or P<0.01). There were different species among the gut microbiota of rats in each group. CONCLUSIONS Ganbao capsules may improve NAFLD by protecting intestinal mucosal barrier function and regulating gut probiotics/harmful bacteria structure.
期刊: 2023年第34卷第08期
作者: 范兴;何艳;杨成梓;林雅婷;黄德福
AUTHORS: FAN Xing,HE Yan,YANG Chengzi,LIN Yating,HUANG Defu
关键字: 肝宝胶囊;非酒精性脂肪性肝病;肠黏膜屏障;肠道菌群;16S核糖体RNA
KEYWORDS: Ganbao capsules; non-alcoholic fatty liver disease; intestinal mucosal barrier; gut microbiota; 16S ribosomal RNA
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