JNK1基因沉默对非酒精性脂肪性肝病小鼠模型高分子量脂联素表达的影响  

Effect on JNK1 gene silencing on the expression of high-molecular-weight adiponectin in mice with nonalcoholic fatty liver disease

作　　者：赵晓芳[1] 唐友明[1] 徐新杰[1] 唐艳芳[1] 刘旭东[1] 吕萍[1] 赵壮志[1] ZHAO Xiaofang;TANG Youming;XU Xinjie(Department of Hepatology,Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530001,China)

机构地区：[1]广西中医药大学附属瑞康医院肝病科

出　　处：《临床肝胆病杂志》2019年第10期2257-2260,共4页Journal of Clinical Hepatology

基　　金：国家自然科学基金(81473532);广西自然科学基金(2016GXNSFAA380251);广西一流学科建设重点课题(2018XK084);广西八桂学者建设专项经费

摘　　要：目的探讨JNK1基因沉默对非酒精性脂肪性肝病(NAFLD)小鼠脂肪组织高分子量脂联素(HMW-APN)及相关通路分子的影响。方法20只C57BL/6小鼠被随机分为正常组、模型对照组、shRNA-JNK1慢病毒处理NAFLD组(JNK1+NAF组)、无关序列shRNA慢病毒处理NAFLD组(无关序列+NAF组),每组5只。正常饮食与高脂饮食分别喂养3个月,成功复制NAFLD小鼠模型,利用最佳干扰效果的shRNA-JNK1慢病毒和无关序列shRNA慢病毒通过尾静脉注射NAFLD小鼠。根据各组饮食及慢病毒注射情况喂养5d后,HE染色观察各组小鼠肝组织的病理学改变。ELISA方法检测各组小鼠血清中HMW-APN的水平。取附睾脂肪垫行WesternBlot检测分析AMPK、P-AMPK、HMW-APN、DsbA-L的表达水平。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果经HE染色,JNK1+NAF组较模型对照组脂滴空泡减少,水肿、炎症减轻。JNK1+NAF组的肝组织脂肪变评分(2.267±0.704)较模型对照组(3.800±0.414)和无关序列+NAF组(3.667±0.617)评分明显降低(P值均<0.05)。经ELISA分析,JNK1+NAF组的血清HMW-APN水平[(294.71±102.30)ng/ml]较模型对照组[(124.06±70.58)ng/ml]明显升高(P<0.001)。经WesternBlot分析,与模型对照组和无关序列+NAF组比较,JNK1+NAF组AMPK、P-AMPK、DsbA-L和HMW-APN表达水平明显升高(P值均<0.05)。结论JNK1基因沉默促进NAFLD小鼠DsbA-L和HMW-APN表达,且JNK1基因沉默可能活化AMPK通路实现对脂联素多聚化调控,从而改善NAFLD脂肪沉积。Objective To investigate the effect of JNK1 gene silencing on the expression of high-molecular-weight adiponectin and related pathway molecules in adipose tissue of mice with nonalcoholic fatty liver disease (NAFLD).Methods A total of 20 C57BL/6 mice were randomly divided into normal group,model control group,NAFLD group treated with shRNA-JNK1 lentivirus (JNK1+NAF group),and NAFLD group treated with unrelated-sequence shRNA lentivirus (unrelated sequence+NAF group),with 5 mice in each group.Normal diet and high-fat diet were given for 3 months,and a mouse model of NAFLD was successfully established.The mice with NAFLD were given tail vein injection of shRNA-JNK1 lentivirus with the optimal interfering effect and unrelated-sequence shRNA lentivirus.After 5 days of feeding based on diet and lentivirus injection,HE staining was used to observe the pathological changes of liver tissue.ELISA was used to measure the serum level of high-molecular-weight adiponectin.Western blot was used to measure the expression of AMP-activated protein kinase (AMPK),phosphorylated AMPK (p-AMPK),high-molecular-weight adiponectin,and disulfide-bond-A oxidoreductase-like protein (DsbA-L) in epididymal fat pad.A one-way analysis of variance was used for comparison of continuous data between groups,and the least significant difference t -test was used for further comparison between two groups.Results HE staining showed that compared with the model control group,the JNK1+NAF group had significant reductions in lipid droplet vacuoles,edema,and inflammation.The JNK1+NAF group had a significantly lower liver steatosis score than the model control group and the unrelated sequence+NAF group (2.267±0.704 vs 3.800±0.414/3.667±0.617,both P <0.05).ELISA showed that the JNK1+NAF group had a significantly higher serum level of high-molecular-weight adiponectin than the model control group (294.71±102.30 ng/ml vs 124.06±70.58 ng/ml,P <0.001).Western Blot showed that the JNK1+NAF group had significantly higher expression levels of AMPK,p-AMPK,DsbA-

关 键 词：非酒精性脂肪性肝病 基因沉默 脂联素 小鼠 近交C57BL 

分 类 号：R575.5[医药卫生—消化系统]