沉默IGFBP7的表达对非酒精性脂肪肝的影响  

Effect of silencing IGFBP7 expression on nonalcoholic fatty liver disease

作　　者：延华[1] 张丹[1] 聂建 李敏[2] YAN Hua;ZHANG Dan;NIE Jian;LI Min(Department of Geriatrics,Shaanxi People′s Hospital,Xi′an 710068,China)

机构地区：[1]陕西省人民医院老年病院,710068 [2]陕西省人民医院保健办,710068

出　　处：《现代消化及介入诊疗》2024年第2期170-175,共6页Modern Interventional Diagnosis and Treatment in Gastroenterology

基　　金：陕西省重点研发项目-社会发展领域(2022SF-450、2021SF-352)。

摘　　要：目的探讨胰岛素样生长因子结合蛋白(insulin-like growth factor binding protein,IGFBP)7在非酒精性病脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)发病中的作用。方法通过喂食高脂饮食(high-fat diet,HFD)建立NAFLD小鼠模型,并通过向肝脏注射腺相关病毒(adeno-associated virus,AAV)介导的短发夹(sh)-IGFBP7敲除IGFBP7。C57BL/6小鼠随机分为4组(每组n=6):正常饮食组(ND)、高脂饮食组(HFD)、HFD空载体注射液组(HFD/AAV-null)、HFD AAV-sh-IGFBP7注射液组(HFD/AAV-sh-IGFBP7)。ND组给与正常饲料12周,HFD组、HFD/AAV-null组、HFD/AAV-sh-IGFBP7组给与高脂饲料4周,之后HFD/AAV-sh-IGFBP7组、HFD/AAV-null组小鼠分别注射2×10^(11) AAV-sh-IGFBP7或AAV-null的基因组拷贝,继续喂食8周。测定各组小鼠体重、肝湿重、肝指数,通过生化分析仪检测各组小鼠血清甘油三酯(TG)、总胆固醇(TC)、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、游离脂肪酸(FFA)、甘油及肝脏TG。采用实时定量PCR、Western blot检测IGFBP7在各组中mRNA,蛋白表达水平及脂肪生成相关的固醇调节元件结合蛋白(sterol regulatory element binding protein,SREBP)1c、脂肪酸合酶(fatty acid synthase,FASN)、乙酰辅酶a羧化酶(acetyl-coa carboxylase,ACC)1和硬脂酰辅酶a去饱和酶(stearoyl coA desaturase,SCD)1的mRNA表达水平。结果与ND组相比,HFD组体重、肝湿重、肝指数及血清中ALT,AST,FFA,TG,TC,甘油、肝TG水平水平显著增加,差异均具有统计学意义(P<0.05),而HFD/AAV-sh-IGFBP7组体重、肝湿重、肝指数及血清中ALT,AST,FFA,TG,TC,甘油、肝TG水平增加均少于HFD/AAV-null组的小鼠,差异均具有统计学意义(P<0.05)。与ND组对比,HFD组小鼠肝脏中IGFBP7mRNA及蛋白的表达水平升高,差异具有统计学意义(P<0.01)。与HFD/AAV-null组相比,HFD/AAV-sh-IGFBP7组小鼠肝脏中IGFBP7蛋白表达水平显著下降,差异具有统计学意义(P<0.01)。与ND组相比,HFD组小鼠肝脏组织中SREBP-1c,FASN,ACC1Objective This study aims to explore the role of Insulin-like growth factor binding protein(IGFBP)7in the pathogenesis of nonalcoholic fatty liver disease(NAFLD).Methods A mouse model of NAFLD was established by feeding a high-fat diet and IGFBP7 was knocked out by injecting adeno-associated virus(AAV)-mediated short hairpin(sh)-IGFBP7 into the liver.The mice were randomly divided into 4 groups(n=6 per group):normal diet group(ND),high fat diet group(HFD),HFD empty carrier injection group(HFD/AAV-null),and HFD AAV-sh-IGFBP7 injection group(HFD/AAV-sh-IGFBP7).ND group was given normal diet for 12 weeks,HFD group,HFD/AAV-null group and HFD/AAV-sh-IGFBP7 group were given high fat diet for 4 weeks.HFD/AAV-sh-IGFBP7 group and HFD/AAV-null group were injected with 2×10^(11) genome copies of AAV-sh-IGFBP7 or AAV-null group,respectively,and then continued to be fed for 8 weeks.Body weight,liver wet weight and liver index of mice in each group were determined.Serum triglyceride(TG),total cholesterol(TC),alanine aminotransferase(ALT),aspartate aminotransferase(AST),free fatty acid(FFA),glycerol and liver TG of mice in each group were detected by biochemical analyzer.Real-time quantitative PCR and Western blot were used to detect IGFBP7 mRNA in each group.Protein expression levels and mRNA expression levels of sterol regulatory element binding protein 1c(SREBP-1c),fatty acid synthase(FASN),acetyl-CoA carboxylase1(ACC1)and stearoyl-CoA desaturase 1(SCD1)related to lipogenesis.Results Compared with ND group,body weight,liver wet weight,liver index and serum ALT,AST,FFA,TG,TC,glycerol and liver TG levels in HFD group were significantly increased,and the differences were statistically significant(P<0.05),while body weight,liver wet weight,liver index and serum ALT,AST,and serum TG levels in HFD/AAV-sh-IGFBP7 group were significantly increased(P<0.05).FFA,TG,TC,glycerol and liver TG levels increased less than those of HFD/AAV-null group,and the differences were statistically significant(P<0.05).Compared with ND group,the expres

关 键 词：NAFLD 高脂饮食 IGFBP7 脂肪变性 脂质代谢 短发卡 

分 类 号：R575.5[医药卫生—消化系统]