摘要
目的:探讨沉默信息调节因子1(SIRT1)过表达介导PPARγ-PGC1α-NRF2通路对肥胖症大鼠肝脏脂肪沉积和变性及氧化应激的影响。方法:首先构建SIRT1慢病毒载体(LV),然后将大鼠随机分为4组:正常对照组(Control)、肥胖症模型组(Obesity model组)、肥胖症+SIRT1对照载体组(Obesity+LV组)和肥胖症+SIRT过表达组(Obesity+LV-SIRT1组)进行后续实验。采用卷尺测量肛鼻长并计算Lee’s指数;罗氏血糖仪检测血糖水平;全自动生化分析仪检测总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平;苏木精-伊红染色法(hematoxylin-eosin staining,HE)和油红O染色检测肝脏脂肪沉积和变性;采用ELISA试剂盒检测总超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)、丙二醛(MDA)和活性氧(ROS)含量;逆转录-聚合酶链反应检测过氧化物酶体增殖物激活受体γ(PPARγ)、PPARγ辅激活子1α(PGC1α)、核因子-E2相关因子2(Nrf2)m RNA水平;蛋白免疫印迹(western blot)检测SIRT1、PPARγ、PGC-1α、Nrf2蛋白表达水平。结果:研究结果表明,与Obesity model组相比,Obesity+LV-SIRT1组SIRT1蛋白水平显著升高(P<0.05);Lee’s指数、体重和血糖水平显著降低(P<0.05);TC、TG、LDL含量显著降低(P<0.05);而HDL含量显著升高(P<0.05);脂肪沉积和变性明显改善;MDA、ROS、LDH含量显著降低(P<0.05);SOD含量显著升高(P<0.05);PPARγ、PGC1α、Nrf2 m RNA和蛋白水平显著升高(P<0.05)。结论:SIRT1过表达可能通过介导PPARγ-PGC1α-NRF2通路对肥胖症大鼠肝脏脂肪沉积和变性及氧化应激起到了调节作用。
Objective:To investigate the effect of silent information regulator 1(SIRT1)overexpression on hepatic lipid accumulation,fatty degeneration,and oxidative stress of liver in obese rats by mediating the PPARγ-PGC1α-NRF2 pathway.Methods:After constructing the SIRT1 lentiviral vector(LV),the rats were divided into control group,obesity model group,obesity+LV group,and obesity+LV+SIRT1 overexpression(obesity+LV-SIRT1)group using a random number table.The nose-anus length was measured by a tape measure and the Lee’s index was calculated;blood glucose level was measured by a Roche blood glucose meter;total cholesterol(TC),triglycerides(TG),low density lipoprotein cholesterol(LDL-C),and high density lipoprotein cholesterol(HDL-C)levels were measured by a chemistry analyzer;hematoxylin-eosin staining(HE)and oil red O were used to detect hepatic lipid accumulation and fatty degeneration of liver;enzyme-linked immunosorbent assay(ELISA)was used to detect superoxide dismutase(SOD),lactate dehydrogenase(LDH),malondialdehyde(MDA)and reactive oxygen species(ROS);reverse transcription-polymerase chain reaction(RT-PCR)was used to detect peroxisome proliferator-activated receptorγ(PPARγ),peroxisome proliferator-activated receptorγcoactivator 1α(PGC1α),and nuclear factor-E2-related factor-2(Nrf2)m RNA levels;Western blot was used to detect SIRT1,PPARγ,PGC-1α,and Nrf2 protein expression levels.Results:Compared with the obesity model group,the results of the study showed that SIRT1 protein level was significantly increased(P<0.05);Lee’s index,body weight,and blood glucose levels were significantly reduced(P<0.05);TC,TG,and LDL were significantly reduced(P<0.05);HDL content was significantly increased(P<0.05);lipid accumulation and fatty degeneration were significantly alleviated;MDA,ROS,and LDH were significantly reduced(P<0.05);SOD was significantly increased(P<0.05);PPARγ,PGC1α,Nrf2 m RNA and protein levels were significantly increased(P<0.05)in the obesity+LV-SIRT1 group.Conclusion:SIRT1 overexpression may mediate the regulation of hepatic lipid accumulation,fatty degeneration,and oxidative stress of the liver in obese rats through the PPARγ-PGC1α-NRF2 pathway.
作者
兰天
刘巍梦
邹阳
邱平
Lan Tian;Liu Weimeng;Zou Yang;Qiu Ping(Department of Endocrinology and Metabolism,the First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China;Department of Gynecology,the Sixth People’s Hospital of Panzhihua City,Sichuan Province,Panzhihua 617023,China;Department of Internal Medicine,the Ninth People’s Hospital of Chongqing,Chongqing 400700,China)
出处
《广西医科大学学报》
CAS
2020年第3期347-355,共9页
Journal of Guangxi Medical University
基金
supported by a grant from the Education Department of Sichuan Province(No.18Z065)。
