胃袖状切除术下调小鼠ChREBP表达改善肝脏脂肪变性  

Down-regulation of ChREBP expression following sleeve gastrectomy to attenuate hepatic steatosis in mice

作　　者：史美龙 杨珵璨 王文越[1] 于洋 刘超凡 王兵[1] SHI Mei-long;YANG Cheng-can;WANG Wen-yue;YU Yang;LIU Chao-fan;WANG Bing(Dept.of General Surgery,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院普外科,上海200011

出　　处：《同济大学学报（医学版）》2021年第3期313-320,共8页Journal of Tongji University(Medical Science)

基　　金：国家自然科学基金(82070917);上海交通大学医学院附属第九人民医院生物样本库项目(YBKB20914)。

摘　　要：目的探讨胃袖状切除术(sleeve gastrectomy,SG)对高糖高脂饮食诱导的肥胖合并非酒精性脂肪肝病(non-alcoholic fatty liver diseases,NAFLD)小鼠肝脏组织中碳水化合物反应元件结合蛋白(carbohydrate responsive element binding protein,ChREBP)表达及肝脏脂肪变性的影响。方法30只4周龄雄性C57BL/6小鼠,随机分为3组:对照(control)组、高糖高脂饮食SG手术(SG)组和高糖高脂饮食假手术(sham)组,每组各10只。喂养20周完成肥胖合并NAFLD小鼠造模。按照SG及sham标准手术方式,对高糖高脂饮食组小鼠分别进行SG手术或sham手术,并记录术后体质量变化。于术后4周和8周分批处死动物,采用H-E、油红染色观察肝脏组织术后病理改变,同时检测肝脏组织中总胆固醇(total cholesterol,TC)、总三酰甘油(total triglycerides,TG)含量,评估小鼠肝脏脂肪变性情况;Western印迹法检测肝脏组织中ChREBP蛋白水平变化。构建ChREBP过表达以及敲低的AML-12细胞,经油酸(oleic acid,OA)诱导后,检测细胞内脂质蓄积情况及TC、TG含量。结果SG术后4周,小鼠体质量较sham组显著下降;小鼠肝脏脂质蓄积面积百分比及肝脏中TC、TG含量及sham组比较,显著改善;肝脏组织中ChREBP的表达及sham组比较,显著下降;SG术后8周,术后疗效较术后4周更为显著。同时体外实验证实细胞内ChREBP的表达上升及下降均可显著改变肝实质细胞内的脂质蓄积面积及TC、TG含量。结论SG手术在术后短期内可有效改善肥胖合并NAFLD小鼠肝脏脂肪变性,其可能与肝脏中ChREBP水平下降有关。Objective To investigate the effects of sleeve gastrectomy(SG)on the expression of carbohydrate responsive element binding protein(ChREBP)and hepatic steatosis in mice.Methods Thirty male C57BL/6 mice of 4-week old were randomly divided into three groups:control group,SG group and sham group with 10 mice in each group.Normal diet was given to mice in control group,while high-sugar-high-fat diet was given to mice in SG and sham groups for 20 weeks to induce non-alcoholic fatty liver diseases(NAFLD),then SG or sham surgery were performed,respectively.Body weight after surgery were recorded.The mice were sacrificed at 4 and 8 weeks after surgery.The pathological changes of liver tissues were observed by H-E and oil red staining,the total cholesterol(TC)and total triglycerides(TG)in liver tissues were measured to assess the hepatic steatosis.Western blotting was performed to detect the changes of ChREBP protein levels in liver tissues.ChREBP overexpressed and knockdown AML-12 cells were constructed and induced by oleic acid(OA)to detect intracellular lipid accumulation and TC and TG contents.Results At 4 weeks after surgery compared with the sham group,the body weight in SG group decreased significantly,the percentage of liver lipid accumulation area and TC and TG contents in the liver improved significantly,the expression of ChREBP in liver tissues decreased significantly.The efficacy was more significant at 8 weeks after SG compared with 4 weeks after SG.In vitro experiments confirmed that the increase or decrease of intracellular ChREBP expression significantly changed the lipid accumulation area and TC and TG contents in hepatocytes.Conclusion SG is effective in improving the degree of hepatic steatosis in obese and NAFLD mice,which might be related to the decreased protein levels of ChREBP in hepatocytes.

关 键 词：胃袖状切除手术 非酒精性脂肪肝 碳水化合物反应元件结合蛋白 小鼠 

分 类 号：R575.5[医药卫生—消化系统]