文献类型：期刊文献

中文题名：半夏泻心汤及其拆方对菌群紊乱幼鼠结肠黏膜免疫的影响

英文题名：Banxia Xiexintang and Its Disassembled Prescriptions Regulate Colonic Mucosal Immunity in Young Rats with Flora Disorder

作者：戴丽蓉[1];陈启明[2];刘喜平[1];林延延[2];李侃[2];岳娟[1];葛艳[2];李沛清[1];朱中博[1];张金铎[2];施丽娟[1]

第一作者：戴丽蓉

机构：[1]甘肃中医药大学甘肃省中药新产品创制工程实验室,甘肃省中医方药挖掘与创新转化重点实验室,兰州730000;[2]兰州大学第一医院,兰州730013

第一机构：甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2022

卷号：28

期号：11

起止页码：42

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金项目(32160255);甘肃省青年科技基金计划项目(21JR7RA388,20JR10RA703,20JR5RA350);甘肃省高等学校创新基金项目(2020B-007);甘肃省委组织部陇原青年创新创业人才项目(2020RCXM183);甘肃省中医方药挖掘与创新转化重点实验室开放基金项目(ZYFY-2020-003)。

语种：中文

中文关键词：半夏泻心汤;肠道菌群;抗生素;肠道免疫;配伍研究

外文关键词：Banxia Xiexintang;intestinal flora;antibiotics;intestinal immunity;study of compatibility

摘要：目的:探讨半夏泻心汤及其拆方对混合抗生素诱导的菌群紊乱幼鼠的治疗作用及可能的作用机制。方法:将70只雄性BALB/c幼鼠随机分为7组,分别为空白组、模型组、双歧杆菌四联活菌片组(西药组,0.68 g·kg^(-1))及半夏泻心汤全方组、辛开组、苦降组、甘补组(9.1、3.19、1.82、4.1 g·kg^(-1)),每组10只。除空白组外,其余各组予混合抗生素灌胃诱导肠道菌群紊乱,14 d后西药组予双歧杆菌四联活菌片灌胃,全方组、辛开组、苦降组及甘补组分别予半夏泻心汤及不同配伍药组灌胃,空白组、模型组给予等体积生理盐水,1次/d,共14 d。连续给药14 d后无菌采集粪便样本用于16SrDNA测序分析肠道菌群,并经腹腔注射脂多糖(LPS,10 mg·kg^(-1))诱导炎性反应,苏木素-伊红(HE)染色观察结肠黏膜组织形态,甲苯胺蓝染色和免疫组织化学观察结肠黏膜巨噬细胞浸润,实时荧光定量聚合酶链式反应(Real-time PCR)检测白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)及白细胞介素-10(IL-10)mRNA的表达情况。结果:与空白组比较,模型组幼鼠肠道微生物结构显著改变(P<0.01),结肠黏膜损伤明显,巨噬细胞浸润减少,炎性因子IL-1β、IL-6、IL-8、TNF-α及IL-10mRNA表达水平均显著降低(P<0.01)。与模型组比较,西药组、半夏泻心汤及其拆方组均能增加菌群相对丰度和物种多样性,提高拟杆菌门、厚壁菌门等有益菌的比例(P<0.01);同时西药组、半夏泻心汤及其拆方组均能改善结肠黏膜损伤(P<0.05,P<0.01),增加巨噬细胞浸润(P<0.05,P<0.01),升高IL-6、IL-8、TNF-αmRNA表达水平(P<0.01);西药组、半夏泻心汤及苦降、甘补组升高IL-1βmRNA表达(P<0.01);西药组、半夏泻心汤及辛开、甘补组IL-10 mRNA表达升高(P<0.05,P<0.01)。结论:半夏泻心汤及拆方组能调整抗生素暴露幼鼠肠道菌群,保护肠道菌群紊乱后结肠黏膜免疫屏障,以半夏泻心汤全方为最佳配伍。
Objective:To explore the therapeutic effect and possible mechanism of Banxia Xiexintang and its disassembled prescriptions in regulating the flora disorder induced by mixed antibiotics in young rats.Method:Seventy male BALB/C young rats were randomly assigned into 7 groups:blank group,model group,Bifidobacterium tetralogy viable tablets(0.68 g·kg^(-1))group,Banxia Xiexintang(9.1 g·kg^(-1))group,Xinkai(3.19 g·kg^(-1))group,Kujiang(1.82 g·kg^(-1))group,and Ganbu(4.1 g·kg^(-1))group,with 10 rats in each group.Except the blank group,the other groups were given mixed antibiotics by gavage to induce intestinal flora disorder.After 14 days,the rats in different drug groups were administrated with corresponding drugs by gavage,and those in the blank group and model group with the same amount of normal saline once a day for 14days.After that,fecal samples were collected aseptically for 16S rDNA sequencing of intestinal flora,and lipopolysaccharide(LPS,10 mg·kg^(-1))was injected intraperitoneally to induce inflammatory reaction.The tissue morphology of colonic mucosa was observed via hematoxylin-eosin(HE)staining,and the macrophage infiltration of colonic mucosa was observed via toluidine blue staining and immunohistochemistry.The expression of interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-8(IL-8),tumor necrosis factor-α(TNF-α),and interleukin-10(IL-10)mRNA were determined by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Result:Compared with the blank group,the modeling changed the intestinal flora structure of the young rats(P<0.01),damaged the colonic mucosa,reduced the macrophage infiltration,and down-regulated the mRNA levels of IL-1β,IL-6,IL-8,TNF-α,and IL-10(P<0.01).Compared with the model group,bifidobacterium quadruple viable tablets,Banxia Xiexintang and its disassembled prescriptions increased the diversity of intestinal flora and the relative abundance of beneficial bacteria such as Bacteroidetes and Firmicutes(P<0.01).At the same time,they ameliorated colonic mucosal injury(P<0.05,P<0.01),increased macrophage infiltration(P<0.05,P<0.01),and up-regulated the mRNA levels of IL-6,IL-8,and TNF-α(P<0.01).The mRNA level of IL-1βwas up-regulated in Bifidobacterium tetralogy viable tablets,Banxia Xiexintang,Kujiang,and Ganbu groups(P<0.01),and that of IL-10 was up-regulated in Bifidobacterium tetralogy viable tablets,Banxia Xiexintang,Xinkai,and Ganbu groups(P<0.01).Conclusion:Banxia Xiexintang and the disassembled prescriptions can adjust the intestinal flora of young rats exposed to antibiotics and protect the immune barrier of colonic mucosa after intestinal flora disorder.In particularly,the whole prescription of Banxia Xiexintang demonstrates the best performance.