白芍总苷长期给药对胶原诱导型关节炎大鼠和正常大鼠肠道菌群影响的纵向研究  

Longitudinal Study on Dynamic Alterations in the Gut Microbiota of Collagen-Induced Arthritis and Normal Rats Following the Prolonged Administration of Total Glucosides of Paeony

作　　者：彭金娥 韩雅馨 卢旭然[2,3] 谢凯莉 南楠 曹炜 龚慕辛[2,3] PENG Jine;HAN Yaxin;LU Xuran;XIE Kaili;NAN Nan;CAO Wei;GONG Muxin(Departmentof Pharmacy,Beijing Ditan Hospital Capital Medical University,Beijing 100015,China;School of TraditionalChinese Medicine,Capital Medical University,Beijing 100069,China;Beijing Key Laboratory of TraditionMedicine Collateral Disease Theory Research,Beijing 100069,China;Guang'anmen Hospital,China Academyof Chinese Medical Sciences,Beijing 100053,China)

机构地区：[1]首都医科大学附属北京地坛医院药学部,北京100015 [2]首都医科大学中医药学院,北京100069 [3]中医络病研究北京市重点实验室,北京100069 [4]中国中医科学院广安门医院,北京100053

出　　处：《中药新药与临床药理》2021年第1期1-16,共16页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金面上项目(81773860)。

摘　　要：目的纵向观察不同剂量白芍总苷(total glucosides of paeony,TGP)给药12周过程中对胶原诱导型关节炎(collagen-induced arthritis,CIA)大鼠以及正常大鼠肠道菌群(简称"菌群")的影响,发现白芍总苷长期给药重点调控的菌群及其代谢功能,以探究其通过影响菌群治疗类风湿关节炎(rheumatoid arthritis,RA)的机制。方法将雄性SD大鼠随机分为正常组、模型组、正常及模型给药白芍总苷(948、474、158 mg·kg^-1)剂量组。于模型复制后给药0周(给药前)以及给药4、8、12周后收集大鼠粪便样本,利用Illumina Miseq平台进行高通量测序,采用相应软件进行操作分类单元(operational taxonomic unit,OTU)、Alpha多样性指数测定,进行Beta多样性分析、基于图形系统发育分析(graphical phylogenetic analysis,GraPhlAn)、偏最小二乘判别分析(partial least squares discriminant analysis, PLS-DA)与线性判别分析(linear discriminant analysis effect size,LEfSe)等的菌群群落结构相似性分析。同时借助数据库进行菌群不同水平分类学组成分析以及PICRUSt(phylogenetic investigation of communities by reconstructing unobserved states)菌群功能的预测。每组先进行不同时间点上述指标的动态变化分析,再进行各组间扣除相同变化后的比较,筛选出白芍总苷对胶原诱导型关节炎和正常大鼠或2个以上剂量调节作用相同的菌群及菌群代谢功能。结果 (1)白芍总苷3个剂量均可影响胶原诱导型关节炎大鼠菌群OTU的经时变化,仅低剂量可影响正常大鼠该指标的经时变化。(2)白芍总苷中、高剂量影响胶原诱导型关节炎大鼠菌群Alpha多样性指数的经时变化,而3个剂量均不影响正常大鼠该指标的经时变化。(3)综合Beta多样性分析以及GraPhlAn、PLS-DA、LEfSe分析,对胶原诱导型关节炎大鼠菌群的影响白芍总苷低、高剂量强于中剂量,对正常大鼠则白芍总苷中、高剂量强于低剂量。Coprococcus属是白芍�Objective In order to find the main gut flora and its metabolic function regulated by total glucosides of paeony(TGP) after long-term administration, and elucidate the mechanism of TGP in the treatment of RA by modulating the flora, we carried out a 12-week longitudinal study in collagen-induced arthritis(CIA) rats and normal rats following the prolonged administration of three doses of TGP(948,474,158 mg·kg^-1).MethodsMale Sprang-Daley(SD)rats were randomly divided into normal group,model group,high,medium and low dose group of TGP. Fecal samples of rats were collected before administration and 4,8 and 12 weeks after administration of TGP. High throughput sequencing was performed by using Illumina Miseq platform. The operational taxon(operational taxonomic unit, OTU) and alpha diversity index were measured with corresponding software, and the similarity analysis of community structure including beta diversity analysis, visualization analysis based on graphical phylogenetic analysis(GraPhlAn), partial least squares discriminant analysis(partial least squares discriminant analysis,PLS-DA)and linear discriminant analysis(linear discriminant analysis effect size,LEfSe)were used. At the same time, the taxonomic composition at different levels and the flora function prediction of PICRUSt(phylogenetic investigation of communities by reconstructing unobserved states) were carried out with the help of database. The dynamic alterations of bacterial community structure and metabolic function of flora were analyzed at different time points in each group. After deductions of the same alterations of flora over time, the comparison between different groups was carried out. The same alterations in the gut antimicrobial caused by TGP administration were screened out in CIA and normal rats groups or in different dose groups.Results(1)All three doses TGP affected the time-dependent changes of OTU in CIA rats, but only low-dose TGP affected the time-dependent changes of OTU in normal rats.(2)The middle and high doses of TGP affect

关 键 词：白芍总苷 胶原诱导型关节炎 肠道菌群 经时变化 菌群代谢功能 大鼠 

分 类 号：R285.5[医药卫生—中药学]