抗纤软肝颗粒通过调节肠道菌群防治肝纤维化

Kangxian Ruangan granules prevents liver fibrosis by regulating the composition of gut microbiota

目的观察抗纤软肝颗粒(KXRG)对肠道菌群的影响,并探究其抗纤维化作用是否与肠道菌群结构变化相关。方法第一部分:药物干预实验。18只C57BL/6小鼠随机分为正常组(n=6)、模型组(n=6)、KXRG组(n=6),40%四氯化碳(CCl_(4))橄榄油溶液sc诱导肝纤维化模型,药物干预8周,无菌操作收集小鼠粪便,并采用16s rRNA测序技术检测小鼠肠道菌群;第二部分:粪菌移植(FMT)验证实验。40只C57BL/6小鼠随机分为对照组(n=10)和模型组(n=30),造模成功后,将第一部分收集的正常组、模型组、KXRG组小鼠粪便,分别移植至新的肝纤维化小鼠模型体内,8周后,检测小鼠体重变化,肝脏指数,血清透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(C-Ⅳ)水平和肝组织形态学改变。结果药物干预实验:与模型组相比,KXRG组肠道菌群在门、科、属等不同水平上均有变化。FMT验证实验:与正常组小鼠粪菌移植组相比,KXRG组小鼠粪菌移植组体重增加量上调,肝脏指数,血清HA、LN、PCⅢ、C-Ⅳ明显下调,肝纤维化病理情况有所改善。结论肠道菌群结构变化在KXRG发挥抗肝纤维化作用中发挥了重要作用。

Objective To observe the effect of Kangxian Ruangan granules(KXRG)on gut microbiota,and to explore whether the anti-liver fibrosis effect of KXRG is related to the composition of gut microbiota.Methods PartⅠ:Experiment of drug intervention.18 C57BL/6 mice are randomly divided into 3 groups:normal group(n=6),model group(n=6)and KXRG group(n=6).The liver fibrosis model is induced by subcutaneous injection of 40%carbon tetrachloride(CCl_(4))olive oil solution,The drug intervention are carried out for 8 weeks,then the feces of mice are collected by aseptic operation,and the structure of gut microbiota of mice is detected by 16S rRNA sequencing method.PartⅡ:Experiment of fecal bacteria transplantation(FMT).40 C57BL/6 mice are randomly divided into 2 groups:control group(n=10),model group(n=30).After being verified,the liver fibrosis mice are treated with the feces of the control group,model group and KXRG group collected in partⅠby intragastric administration.8 weeks later,body weight growth,liver index,serum hyaluronic acid(HA),layer adhesion protein(LN),typeⅢbefore collagenⅢ(PCⅢ),typeⅣcollagen(IV-C)level and liver tissue morphology change are detected.Results Experiment of drug intervention:Compared with model group,the composition of gut microbiota in KXRG group are changed at phylum,family and genus level.Experiment of FMT:Compared with model FMT group,the expression of KXRG FMT group on body weight growth is up-regulated,the expression on liver index,serum HA,LN、PCⅢ、C-Ⅳlevel are down-regulated,the pathological and morphological changes of liver are improved significantly,Conclusion The composition of gut microbiota may play a crucial role in the anti-fibrosis effect of KXRG.