葛根芩连汤干预胰岛素抵抗大鼠的粪便代谢组及相关菌群研究  

Study on Fecal Metabolic Mechanism and Related Flora of Gegen Qin Lian Decoction in Rats with Insulin Resistance

作　　者：彭程程 姚亮亮[1] 曾国威 曾治君[1,2,3] 盛译萱 李冰涛 张启云[1,2,3] 姜丽 刘海云[1] 徐国良 PENG Chengcheng;YAO Liangliang;ZENG Guowei;ZENG Zhijun;SHENG Yixuan;LI Bingtao;ZHANG Qiyun;JIANG Li;LIU Haiyun;XU Guoliang(Research Center for Differentiation and Development of Traditional Chinese Medicine(TCM)Basic Theory,Jiangxi University of TCM,Nanchang 330004 Jiangxi,China;Jiangxi Key Laboratory of TCM Etiopathogenesis,Nanchang 330004 Jiangxi,China;Jiangxi Key Laboratory of Pharmacology of TCM,Nanchang 330004 Jiangxi,China;Guangzhou First People's Hospital,Guangzhou 510180 Guangdong,China)

机构地区：[1]江西中医药大学中医基础理论分化发展研究中心,江西南昌330004 [2]江西省中医病因生物学重点实验室,江西南昌330004 [3]江西省中药药理重点实验室,江西南昌330004 [4]广州市第一人民医院,广东广州510180

出　　处：《中药新药与临床药理》2021年第12期1737-1744,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81560744,81760787);江西省自然科学基金项目(20192BAB205094);中药学一流学科建设科研启动基金项目(JXSYLXK-ZHYAO117);江西省教育厅科学技术研究项目(GJJ190643)。

摘　　要：目的从代谢组学及肠道菌群角度探讨葛根芩连汤干预胰岛素抵抗(IR)大鼠肠道代谢紊乱的作用机制。方法将雄性SD大鼠随机分成正常组、模型组及葛根芩连汤低、中、高剂量(1.65、4.96、14.86 g·kg-1)组。采用高脂饲料喂养12周建立IR大鼠模型;模型复制成功后,连续灌胃给药16周,每天1次,同时继续以高脂饲料喂养。给药结束后,采用超高效液相色谱-串联四级杆-飞行时间质谱(UHPLC/Q-TOF-MS)结合多元分析的代谢组学技术对大鼠的粪便样品进行代谢轮廓分析,分析各组大鼠粪便内源性代谢物的变化,鉴定潜在的生物标记物并进行生物学意义分析。对粪便样品进行16S rRNA V3~V4区域测序,通过两部分模型(Twopart model)对生物标记物与肠道菌群进行关联性分析。结果筛选和鉴定出5个潜在生物标记物:咪唑丙酸、吲哚丙烯酸、二十碳四烯酸、二十四碳六烯酸及十二烷二酸;与模型组比较,葛根芩连汤低剂量组的十二烷二酸含量明显增加(P<0.05),二十四碳六烯酸含量明显减少(P<0.05)。肠道菌群关联分析结果显示,二十碳四烯酸和十二烷二酸共关联到5个肠道菌群操作分类单元(OTU),涉及到的肠道菌群为厚壁菌门和拟杆菌门,其中厚壁菌门消化链球菌科Romboutsia属为二者所共有,其余OTU为十二烷二酸特有;另外3个生物标记物未关联到相关肠道菌群。结论葛根芩连汤对高脂诱导的IR大鼠的预防作用可能与其改善肠道中内源性代谢物及相关肠道菌群的水平,恢复体内正常代谢活动有关。Objective To investigate the mechanism of Gegen Qin Lian decoction on intestinal metabolic disorder in insulin resistance(IR)rats from the perspectives of metabonomics and intestinal microflora.Methods Male SD rats were randomly divided into normal control group,model group and low,medium and high dose groups of Gegen Qin Lian decoction(1.65,4.96,14.86 g·kg-1).IR model was established by feeding high fat for 12 weeks.After the model was successfully replicated,drugs were administered intragastrically once a day for 16 consecutive weeks,while the high fat diet was continued.After administration,the fecal profiles of rats were analyzed by UHPLC-QTOF-MS,the changes of endogenous metabolites in feces of rats in each group were analyzed to identify potential biomarkers and explore their biological significance.DNA of fecal samples were sequenced at 16 S rRNA V3-V4 region,and the Two-part model was used to analyze relationship between biomarkers and intestinal flora.Results After screening,5 potential biomarkers including imidazole propionic acid,indole acrylic acid,eicosatetraenoic acid,docosahexaenoic acid,and dodecanedioic acid,were identified.Compared with IR group,the content of dodecanedioic acid in the low-dose Gegen Qin Lian decoction group was significantly increased(P<0.05),and the content of eicosapentaenoic acid was significantly decreased(P<0.05).The association analysis of gut microbiota showed that eicosatetraenoic acid and dodecanedioic acid were associated with five operating taxonomic units(OTU)of gut microbiota belonged to Firmicutes and Bacteroidetes;of which the Peptostreptococcaceae(Romboutsia)that belonged to Firmicutes is shared by both and the remaining OTUs were endemic to the dodecanedioic acid.The other3 biomarkers were not associated with the relevant intestinal microflora.Conclusion The preventive effect of Gegen Qin Lian decoction on high-fat-induced IR rats may function by improving the levels of endogenous metabolites and regulating related intestinal flora in the intestine,thereby res

关 键 词：胰岛素抵抗 葛根芩连汤 代谢组学 机制 肠道菌群 大鼠 

分 类 号：R285.5[医药卫生—中药学]