丁酸梭菌对小鼠非酒精性脂肪性肝病的改善作用及对细胞程序性坏死的影响  

Ameliorative effect of Clostridiun butyricum on non-alcoholic fatty liver disease in mice and its effect on necroptosis

作　　者：赵翠娟 陈烨 李妍华 武文博 陈强 ZHAO Cuijuan;LI Yanhua;WU Wenbo;CHEN Ye;CHEN Qiang(Department of Gastroenterology,Inner Mongolia Baogang Hospital,Third Affiliated Hospital of Inner Mongolia Medical University,Baotou 014000,China;Students’Affairs Office,Baotou Medical College,Baotou 014000,China;Department of Plastic Surgery,Fourth Hospital of Baotou,Baotou 014000,China)

机构地区：[1]内蒙古包钢医院内蒙古医科大学第三临床医学院消化内科,内蒙古包头014000 [2]包头医学院学生处,内蒙古包头014000 [3]包头市第四医院整形外科,内蒙古包头014000

出　　处：《国际消化病杂志》2025年第2期87-93,共7页International Journal of Digestive Diseases

基　　金：内蒙古自治区科技计划项目(2022YFSH0042);航天医疗健康科技集团有限公司科研项目(2023YK22);内蒙古医学科学院公立医院科研联合基金科技项目(2023GLLH0242)。

摘　　要：目的探讨丁酸梭菌(CB)对小鼠非酒精性脂肪性肝病(NAFLD)的改善作用及对细胞程序性坏死的影响。方法选取C57BL/6小鼠随机分为正常饮食组(NC组)、高脂饮食组(HFD组)和CB干预组(CB组),每组6只。NC组采用正常饲料喂养12周,HFD组和CB组均采用高脂饲料喂养12周以建立NAFLD动物模型。自实验第6周起,CB组每日给予CB 0.2 mL灌胃,NC组和HFD组均每日给予0.9%氯化钠溶液0.2 mL灌胃。每周称量并记录小鼠的体质量。第12周后,所有小鼠均采血后处死,称量肝脏质量并计算肝脏指数。采用ELISA法检测3组的血清ALT、AST水平。采用H-E染色法观察小鼠肝脏病理变化。采用油红O脂肪染色法观察小鼠肝内脂肪沉积情况。采用实时荧光定量PCR法检测3组的肝组织中受体相互作用蛋白激酶3(RIPK3)、混合谱系激酶结构域样蛋白(MLKL)、TNF-α、IL-1β、IL-6 mRNA表达水平,采用蛋白质印迹法检测3组的肝组织中RIPK3、MLKL蛋白表达水平,评估3组的肝脏细胞程序性坏死情况。结果与NC组相比较,HFD组的体质量和肝脏指数均显著增高(P均<0.001),血清ALT、AST水平均显著升高(P均<0.001),有明显的肝细胞内脂肪沉积,肝组织中TNF-α、IL-1β、IL-6 mRNA表达水平均显著升高(P均<0.01),肝组织中RIPK3、MLKL的mRNA和蛋白表达水平均显著升高(P均<0.01)。与HFD组相比较,CB组的体质量和肝脏指数均显著降低(P均<0.05),血清ALT、AST水平均显著降低(P均<0.001),肝内脂肪沉积有显著改善,肝组织中TNF-α、IL-1β、IL-6 mRNA表达水平均显著降低(P均<0.01),肝组织中RIPK3、MLKL的mRNA和蛋白表达均显著降低(P均<0.05)。结论CB可以减轻NAFLD小鼠的肝内脂肪沉积和炎症反应,该作用与其抑制RIPK3、MLKL表达,从而抑制肝脏细胞程序性坏死相关。Objective This study aims to explore the ameliorative effect of Clostridium butyricum(CB)on non-alcoholic fatty liver disease(NAFLD)in mice and its effect on necroptosis.Methods C57BL/6 mice were randomly divided into three groups:a normal diet group(NC group),a high-fat diet group(HFD group),and a CB intervention group(CB group),with 6 mice in each group.The NC group was fed with a normal diet for 12 weeks,while both the HFD group and CB groups were fed with a high-fat diet to establish a NAFLD animal model.From the 6th week,the CB group was given 0.2 mL of CB daily via gavage,while the NC and HFD groups were given 0.2 mL of 0.9%sodium chloride solution daily.Body weight was recorded weekly.After 12 weeks,all mice were euthanized after blood collection.Liver mass was measured,liver-index calculated.ELISA method was used to detect the serum ALT and AST levels of the three groups.H-E staining was performed to observe liver pathology.Oil Red O staining was used to observe the lipid accumulation in mice liver.Real-time fluorescence quantitative PCR was used to quantify the mRNA expression of receptor interacting protein kinase 3(RIPK3),mixed lineage kinase domain-like protein(MLKL),TNF-α,IL-1β,and IL-6 in the liver tissue.Western blotting was used to detect the protein expression of RIPK3 and MLKL in the liver tissue to evaluate the necroptosis of liver cells.Results Compared with the NC group,the HFD group shows significantly increased body weight and liver index,elevated serum ALT and AST levels(P<0.001),and noticeable lipid deposition in hepatocytes.The mRNA expression of TNF-α,IL-1β,and IL-6 in liver tissue,along with RIPK3 and MLKL mRNA and protein levels,are significantly higher in the HFD group(P<0.01).In contrast,the CB group show significantly reductions in body weight,liver index(P<0.05),serum ALT and AST levels(P<0.001),and lipid deposition in hepatocytes.The mRNA expression of TNF-α,IL-1β,and IL-6 in liver tissue(P<0.01),as well as RIPK3 and MLKL and protein expression(P<0.05),are significantly red

关 键 词：非酒精性脂肪性肝病 丁酸梭菌 程序性坏死 炎症反应 

分 类 号：R575.5[医药卫生—消化系统]