基于16S rDNA测序技术分析益气活血方对脑缺血损伤大鼠肠道菌群多样性的影响  

Analysis of the Effect of Yiqi Huoxue Prescription on the Diversity of Intestinal Flora in Rats with Cerebral Ischemia Injury Based on 16S rDNA Sequencing Technology

作　　者：瞿光友 彭友飞 陈怡名 王月 王连芝[1] 张淑香[1] 顾媛媛[1] QU Guangyou;PENG Youfei;CHEN Yiming;WANG Yue;WANG Lianzhi;ZHANG Shuxiang;GU Yuanyuan(Institute of Traditional Chinese Medicine,Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学中医药研究院,黑龙江哈尔滨150040

出　　处：《现代中药研究与实践》2024年第4期24-30,共7页Research and Practice on Chinese Medicines

基　　金：黑龙江中医药大学科研基金(2017bs02);教育部重点实验室-北药基础与应用研究开放课题基金;哈尔滨市科技局科技创新人才应用技术研究与开发(青年后备人才A类)项目(2017RAQXJ127);黑龙江省教育厅创新人才支持计划(UNPYSCT-2017218)。

摘　　要：目的 基于16S rDNA测序技术研究益气活血方剂对脑缺血损伤大鼠肠道菌群多样性的影响,从“脑-肠”轴角度探讨该方剂抗脑缺血损伤的作用机制。方法 将SD雄性大鼠36只随机分为6组,每组6只,即假手术组、模型组、尼莫地平组、益气活血方高、中、低剂量组。连续灌胃14 d后,采用结扎双侧颈总动脉法建立急性脑缺血损伤模型,苏木精-伊红(HE)染色观察脑组织病理学变化,ELISA法检测大鼠血清中炎性因子IL-6和TNF-α的含量。收集各组大鼠结肠内容物,16S rDNA测序法检测粪便肠道菌群的变化。结果 HE染色观察结果显示益气活血方不同剂量组对脑缺血损伤程度有明显改善作用,且存在量效相关性。与假手术组比较,模型组大鼠血清中炎性因子 IL-6、TNF-α的含量明显升高(P<0.05);益气活血方高剂量组干预后降低(P<0.05,P<0.01)。16S rDNA测序结果显示,益气活血方可调节脑缺血损伤大鼠肠道菌群结构,增加大鼠Chao1、Observed OTUs、Simpson和Shannon指数,Beta多样性分析结果表明所有样本可明显分为三组,聚类良好。益气活血方可上调厚壁菌门(Firmicutes)、髌骨细菌门(Patescibacteria)的丰度(P<0.05),下调疣微菌门(Verrucomicrobia)的丰度(P<0.05);上调Lachnospiraceae_NK4A136_group、Firmicutes_unclassified、Alloprevotella的相对丰度(P<0.05),下调拟杆菌属(Bacteroides)、阿克曼氏菌(Akkermansia)、梭菌属(Clostridium_sensu_stricto_1)、Romboutsia、Veillonella、双歧杆菌属(Bifidobacterium)的丰度(P<0.05)。结论 益气活血方对脑缺血损伤大鼠脑组织具有保护作用,其作用机制可能与降低血清炎症,调节肠道菌群微生物组成、结构及群落丰度有关,进一步验证了“脑-肠”轴学说。Objective To investigate the effects of Yiqi Huoxue prescription on the diversity of intestinal flora in rats with cerebral ischaemia injury based on 16S rDNA sequencing technology and explore the mechanism of anti-ischemic injury of the formula from the perspective of the"brain-gut"axis.Methods 36 male SD rats were randomly divided into 6 groups,with 6 rats in each group,namely sham operation group,model group,nimodipine group and high,medium and low dose groups of Yiqi Huoxue prescription.After 14 d of continuous gavage,a model of acute cerebral ischaemic injury was established using the bilateral common carotid artery ligation method.The pathological changes of brain tissue were observed by hematoxylin-eosin(HE)staining,and IL-6 and TNF-αin serum were detected by ELISA.Colonic contents were collected from each group of rats,and changes in faecal intestinal flora were detected by 16S rDNA sequencing.Results Different dosage groups of Yiqi Huoxue prescription could obviously improve the degree of cerebral ischemia injury,and there was a dose-effect correlation.Compared with the sham operation group,the serum levels of IL-6 and TNF-αin the model group increased significantly(P<0.05).Compared with the sham-operated group,the serum levels of the inflammatory factors IL-6 and TNF-αwere significantly higher in the model group of rats(P<0.05);they were significantly lower in the high-dose group of Yiqi Huoxue prescription after the intervention(P<0.05,P<0.01).The results of 16S rDNA sequencing showed that Yiqi Huoxue prescription could regulate the structure of intestinal flora in rats with cerebral ischaemia injury,and increase Chaol,Observed OTUs,Simpson and Shannon indices in rats,and the results of Beta Diversity Analysis showed that all the samples could be obviously divided into three groups with good clustering.Yiqi Huoxue prescription could significantly increase the abundance of Firmicutes and Patescibacteria(P<0.05),significantly decreased the abundance of Verrucomicrobia(P<0.05),and significantly increas

关 键 词：益气活血方 脑缺血损伤 肠道菌群 16S rDNA测序 

分 类 号：R285.5[医药卫生—中药学]