矿物药青礞石对戊四唑点燃癫痫大鼠肠道菌群的影响  

Effects of mineral Chinese medicine Chloriti Lapis on gut microbiota in PTZ-kindled epileptic rats

作　　者：袁鹏 马瑜璐 刘圣金 房方 杨文国 卞勇 徐晨昱 张志杰[2] 奥•乌力吉 段金廒 YUAN Peng;MA Yu-lu;LIU Sheng-jin;FANG Fang;YANG Wen-guo;BIAN Yong;XU Chen-yu;ZHANG Zhi-jie;AO·Wuliji;DUAN Jin-ao(State Administration of Traditional Chinese Medicine Key Laboratory of Chinese Medicinal Resources Recycling Utilization,National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine,Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Inner Mongolia University for Nationalities,Tongliao 028000,China)

机构地区：[1]南京中医药大学药学院/江苏省中药资源产业化过程协同创新中心/中药资源产业化与方剂创新药物国家地方联合工程研究中心/国家中医药管理局中药资源循环利用重点研究室,江苏南京210023 [2]中国中医科学院中药研究所,北京100700 [3]内蒙古民族大学,内蒙古通辽028000

出　　处：《中草药》2021年第7期2011-2023,共13页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(81673566);全国矿物药资源普查项目(2019);江苏省研究生科研与实践创新计划项目(KYCX18-1608);2017年中医药公共卫生服务补助专项“全国中药资源普查项目”(财社[2017]66号)。

摘　　要：目的研究青礞石对戊四唑(pentylenetetrazol,PTZ)点燃癫痫大鼠肠道菌群的影响,从肠道微生态环境视角探讨青礞石治疗癫痫疾病可能的作用机制。方法以青礞石粉末为研究对象,用PTZ点燃法建立大鼠癫痫动物模型,实验分为对照组、模型组、卡马西平组和青礞石组(高剂量组为临床用量的20倍、低剂量组为临床用量的5倍)。给药4周后,取各组大鼠结肠内容物,通过16Sr RNA测序对各组大鼠肠道菌群的多样性进行分析,利用UPARSE、SPSS18.0等对结果进行生物信息学及统计学分析。结果α多样性分析可知,模型组大鼠肠道菌群的Chao 1、Ace丰富度指数与对照组相比没有显著性变化;与对照组、模型组相比,青礞石组尤其是高剂量组对细菌群落丰富度有显著影响(P<0.05)。Shannon、Simpson群落多样性指数分析结果显示,模型组与对照组无显著性差异;青礞石组对群落多样性干预不明显。青礞石主要干预肠道菌群的丰富度、卡马西平主要干预肠道菌群的多样性。β多样性各组层级聚类均能较好地分开,其中青礞石高剂量组与低剂量组部分样本不能明显分开,2组样本可归为一类,对照组、模型组可归为一类,卡马西平组单独归为一类。3D-主成分分析(3D-principal component analysis,3D-PCA)、3D-主坐标分析(3D-principal co-ordinates analysis,3D-PCoA)结果显示,各组菌群结构轮廓均能明显分开,青礞石低剂量组介于对照组和模型组之间,菌群结构轮廓部分与模型组、对照组有重合交叉,且有偏向对照组的明显趋势。非度量多维尺度(nonmetric multidimensional scaling,NMDS)分析结果显示,对照组、模型组、青礞石组集中在一个区域,青礞石组与模型组、对照组有重合交叉,青礞石高剂量组、低剂量组对肠道菌群结构有相似的干预效果。物种组成门水平分析可知,各组大鼠的优势菌群主要有厚壁菌门(Firmicutes)、拟杆菌门(BacteroideteObjective To study the effect of Chloriti Lapis on the gut microbiota in pentylenetetrazol(PTZ)-kindled epileptic rats, and explore the possible mechanism of Chloriti Lapis in the treatment of epilepsy from the perspective of intestinal micro-ecological environment. Methods Taking Chloriti Lapis powder as the research object, the rat epilepsy animal models were established by the PTZ kindling method. The experiment was divided into blank group, model group, carbamazepine group, and Chloriti Lapis group(the high-dose group was 20 times of the clinical drug, the low-dose group was five times). After 4 weeks of treatment, the colon contents of each group of rats were taken, and the diversity of the gut microbiota of each group of rats were analyzed by 16 S rRNA sequencing, and the results were analyzed in bioinformatics and statistics using UPARSE, SPSS 18.0, etc. Results The α-diversity analysis showed that, compared with the blank group, the Chao 1 and Ace richness index of the gut microbiota flora in the model group had no significant change;Compared with the blank group and the model group, the Chloriti Lapis group, especially the high-dose group, had a significant impact on the richness of gut microbiota flora(P<0.05). The results of Shannon and Simpson index analysis showed that there was no significant difference between the model group and the blank group. Chloriti Lapis mainly interfered with the richness of gut microbiota flora, and carbamazepine mainly interfered with the diversity of gut flora. The results showed that the hierarchical clustering of each group of β-diversity could be well separated. Some of the samples in Chloriti Lapis high-dose group and low-dose group could not be separated obviously, and the two groups of samples could be classified into one group. The blank group and the model group could be classified into one group, and the carbamazepine group could be classified into one group. The results of 3 D-PCA and 3 D-PCoA analysis showed that the bacterial structure outline of each group

关 键 词：矿物药 青礞石 癫痫 戊四氮 结肠 肠道菌群 脑-肠轴 作用机制 

分 类 号：R285[医药卫生—中药学]