摘要
目的:探究茵陈四苓颗粒联合利福昔明对急性肝衰竭(ALF)小鼠肠道菌群及肠屏障功能的影响。方法:50只小鼠随机分为对照组(Control组)、模型组(ALF组)、茵陈四苓颗粒组(YCSLG组)、利福昔明组(RIF组)和茵陈四苓颗粒联合利福昔明组(YCSLG+RIF组),每组10只。采用腹腔注射2次硫代乙酰胺(TAA,300 mg/kg)建立ALF小鼠模型。YCSLG组、RIF组、YCSLG+RIF组小鼠以茵陈四苓颗粒(1.5 g/kg)和利福昔明(15 mg/kg)灌胃,Control组、ALF组小鼠以等量0.9%氯化钠溶液灌胃,早晚各灌胃1次,连续3 d。造模成功48 h后,收集腹主动脉血,检测小鼠血清肝功能指标[丙氨酸氨基转移酶(ALT)、天冬氨酸基转移酶(AST)、总胆红素(TBIL)]及肠黏膜损伤相关蛋白[二胺氧化酶(DAO)、D-乳酸(D-LA)]水平;ELISA检测血清炎症因子IL-1β、IL-6、TNF-α水平;HE染色观察小鼠肝组织和结肠组织病理损伤;免疫组化染色、Western blot检测结肠组织肠黏膜愈合相关蛋白[紧密连接蛋白4(Claudin-4)、封闭蛋白(Occludin)、闭锁小带蛋白1(ZO-1)]水平;16S rRNA测序检测小鼠粪便中肠道菌群结构变化。结果:与Control组相比,ALF组小鼠血清ALT、AST、TBIL、IL-1β、IL-6、TNF-α、DAO、D-LA水平升高,结肠组织Claudin-4、Occludin、ZO-1蛋白水平降低,粪便中乳酸杆菌属(Lactobacillus)、杜氏菌属(Dubosiella)、普雷沃氏菌属(Prevotella)、阿克曼菌属(Akkermansia)、粪杆菌属(Faecalibaculum)等益生菌丰度降低,志贺氏菌属(Shigella)、苏黎世杆菌属(Turicibacter)、克雷伯氏菌(Klebsiella)、Romboutsia、拟杆菌属(Bacteroides)等致病菌丰度增加(P<0.05);与ALF组相比,YCSLG组、RIF组、YCSLG+RIF组小鼠血清ALT、AST、TBIL、IL-1β、IL-6、TNF-α、DAO、D-LA水平降低,结肠组织Claudin-4、Occludin、ZO-1蛋白水平升高,粪便中Lactobacillus、Dubosiella、Prevotella、Akkermansia、Faecalibaculum等益生菌丰度增加;Shigella、Turici-bacter、Klebsiella、Romboutsia、Bacteroides等致病菌丰度降低(P<0.05);且YCSLG+RIF组的治疗效果优于YCSLG组和RIF组(P<0.05)。结论:茵陈四苓颗粒联合利福昔明能够提高ALF小鼠肝功能,改善肠道菌群结构,并恢复肠屏障功能。
Objective:To explore the effect of Yinchensiling granules combined with rifaximin on the gut microbiota and intes-tinal barrier function in mice with acute liver failure(ALF).Methods:Fifty mice were randomly divided into Control group,model group(ALF group),Yinchensiling granules group(YCSLG group),Rifaximin group(RIF group)and Yinchensiling granules com-bined with Rifaximin group(YCSLG+RIF group),with ten mice in each group.ALF mice model were established by intraperitoneal injection of thioacetamide(TAA,300 mg/kg)twice.YCSLG group,RIF group and YCSLG+RIF group mice were orally administered Yinchensiling granules(1.5 g/kg)and rifaximin(15 mg/kg),while Control group and ALF group mice were orally administered an equal amount of 0.9%sodium chloride solution,gavage once in the morning and once in the evening for 3 consecutive days.48 hours after successful modeling,abdominal aortic blood was collected to detect liver function indicator[alanine aminotransferase(ALT),as-partate aminotransferase(AST),total bilirubin(TBIL)]levels and intestinal mucosal injury related protein[diamine oxidase(DAO),D-lactic acid(D-LA)]in mice serum;ELISA was used to detect inflammatory factor IL-1β,IL-6,TNF-αlevels in serum;HE staining was used to observe pathological damage in liver and colon tissues.Immunohistochemical staining and Western blot were used to detect intestinal mucosal healing related protein[Claudin-4,Occludin,zonula occludens 1(ZO-1)]levels in colon tissue;16S rRNA sequencing was used to detect changes in gut microbiota structure in mouse feces.Results:Compared with Control group,serum ALT,AST,TBIL,IL-1β,IL-6,TNF-α,DAO and D-LA levels in ALF group were increased,Claudin-4,Occludin,ZO-1 pro-tein levels in colon tissue were decreased,the abundance of probiotics such as Lactobacillus,Dubosella,Prevotella,Akkermansia and Faecalibacterium were decreased,while the abundance of pathogenic bacteria such as Shigella,Zurich,Klebsiella,Romboutsia and Bacteroides were increased(P<0.05);compared with ALF group,serum ALT,AST,TBIL,IL-1β,IL-6,TNF-α,DAO and D-LA levels in YCSLG group,RIF group and YCSLG+RIF group were decreased,Claudin-4,Occludin,ZO-1 protein levels in colon tissue were increased,the abundance of probiotics such as Lactobacillus,Dubosella,Prevotella,Akkermansia and Faecalibacterium in feces were increased,while the abundance of pathogenic bacteria such as Shigella,Zurich,Klebsiella,Romboutsia and Bacteroides were de-creased(P<0.05);and the treatment effect of YCSLG+RIF group was better than that of YCSLG group and RIF group(P<0.05).Con-clusion:Yinchensiling granules combined with rifaximin can improve liver function,improve intestinal microbiota structure,and re-store intestinal barrier function in ALF mice.
作者
吕琳
王京
张琪瑶
LYU Lin;WANG Jing;ZHANG Qiyao(Health Care Department of Guang'anmen Hospital Jinan Hospital(Jinan Hospital of Traditional Chinese Medicine),Jinan 250012,China;Department of Infectious Diseases,Shandong Provincial Hospital Affiliated to Shandong First Medical University,Jinan 250021,China;Depart-ment of Pharmacy,Jinan Hospital,Guang'anmen Hospital(Jinan Hospital of Traditional Chinese Medicine),Jinan 250012,China)
出处
《中国免疫学杂志》
北大核心
2025年第8期1833-1839,共7页
Chinese Journal of Immunology
基金
山东省自然科学基金(ZR2021MH068)。
关键词
急性肝衰竭
茵陈四苓颗粒
利福昔明
肠道菌群
肠屏障功能
Acute liver failure
Yinchensiling granules
Rifaximin
Intestinal microbiota
Intestinal barrier function
