菌黄保肠合剂对脓毒症大鼠肠道微生态调节作用的影响  

Effect of Junhuang Baochang Mixture on the regulation of intestinal microecology in septic rats

作　　者：铁明慧 陈斌 庞永诚 李巽华 刘明 陈维军 TIE Minghui;CHEN Bin;PANG Yongcheng;LI Xunhua;LIU Ming;CHEN Weijun(Department of Critical Care Medicine,the Third Affiliated Hospital of Yunnan University of Chinese Medicine,Kunming,Yunnan 650500)

机构地区：[1]云南中医药大学第三附属医院重症医学科,云南昆明650500

出　　处：《河北中医》2023年第7期1140-1146,共7页Hebei Journal of Traditional Chinese Medicine

基　　金：云南省科技厅应用基础研究联合专项[编号:2018FF001-(025)]。

摘　　要：目的研究探讨菌黄保肠合剂对脓毒症大鼠肠道微生态的调节作用。方法将80只雄性SD大鼠随机分为假手术组、模型组、菌黄保肠合剂组及双歧杆菌组,每组20只。假手术组仅开腹分离并暴露盲肠,其余各组开腹后进行盲肠结扎穿刺(CLP)手术,术后24 h后切除坏死盲肠并给予盐酸头孢噻呋钠皮下注射建立脓毒症致肠道微生态变化模型,术后48 h各组开始给予相关药物灌胃。菌黄保肠合剂组按4.0 g/(kg·d)予菌黄保肠合剂灌胃,双歧杆菌组按0.1 g/(kg·d)予双歧杆菌活菌胶囊药液灌胃,假手术组和模型组予3 mL纯净水灌胃,共7天,记录给药7天内大鼠存活率。灌胃结束后处死大鼠,收集大鼠结直肠内粪便标本,利用Illumina Hiseq高通量测序技术进行大鼠的肠道菌群宏基因多样性分析,计算细菌群落在门分类学相对丰度、Alpha多样性指数,分析beta多样性,使用R软件绘制出主坐标成分分析图(PCoA),并采用在线分析程序LEfSe寻找组间在丰度上有显著差异的物种。结果①7天存活率比较:假手术组存活率100%,模型组存活率55%,菌黄保肠合剂组存活率70%,双歧杆菌组存活率65%,与假手术组比较,模型组、菌黄保肠合剂组、双歧杆菌组大鼠存活率显著降低(P<0.05)。②细菌群落在门分类学相对丰度比较:与假手术组比较,模型组拟杆菌门、变形杆菌门相对丰度明显上调(P<0.05),厚壁菌门、放线菌门相对丰度明显下调(P<0.05);与模型组比较,菌黄保肠合剂组厚壁菌门相对丰度明显上调(P<0.05),拟杆菌门、变形杆菌门相对丰度明显下调(P<0.05),双歧杆菌组仅变形杆菌门相对丰度明显下调(P<0.05);与双歧杆菌组比较,菌黄保肠合剂组拟杆菌门相对丰度明显下调(P<0.05)。③肠道微生态alpha多样性分析:与假手术组比较,模型组Chao1指数和PD_whole_tree指数均下降(P<0.05);与模型组比较,菌黄保肠合剂组及双歧杆菌组Chao1指数和PD_wholeObjective To investigate the regulatory effect of Junhuang Baochang Mixture(JHBC)on intestinal microecology in septic rats.Methods Eighty male SD rats were randomly divided into four groups,including:sham operation group,model group,JHBC group and bifidobacterium group according to the body weight(n=20).In the sham operation group,only the cecum was separated and exposed by laparotomy.The remaining sixty rats in the other three groups underwent cecal ligation and puncture(CLP)after laparotomy.The necrotic cecum was removed 24 hours after surgery,and ceftiofur sodium hydrochloride was injected subcutaneously to establish a sepsis-induced intestinal microecological change model.The related drugs were given by gavage to each group 48 hours after surgery,for a total of 7 days.In the JHBC group,4.0 g·(kg·d)-1 JHBC was administered by gavage;in the bifidobacterium group,0.1 g·(kg·d)-1 bifidobacterium viable capsules was administered by gavage;in sham operation and model groups,3 mL of purified water was administered by gavage.After the gavage,the rats were sacrificed,and fecal specimens were collected from colon and rectum.The survival rate of rats within 7 days of administration was recorded.The Illumina Hiseq high-throughput sequencing technology was used to analyze the diversity of the intestinal flora structure of rats.The relative abundance of the bacterial community in the phylum taxonomy and alpha diversity index were calculated,and the beta diversity was analyzed.R software was applied to draw the principal coordinate component analysis diagram(PCoA),and the online analysis program linear discriminant analysis effect size(LEfSe)was used to search for species with significant differences in abundance between groups.Results(1)7-day survival rate:100%in the sham operation group,55%in the model group,70%in the JHBC group and 65%in the bifidobacterium group.Compared with the sham operation group,the survival rate in the model group,the JHBC group and the bifidobacterium group was significantly reduced(P<0.05).(2

关 键 词：胃肠道微生物组 动物实验 大鼠 脓毒症 中药疗法 

分 类 号：R285.5[医药卫生—中药学]