PNPLA3基因高表达对非酒精性脂肪性肝病模型细胞增殖和甘油三酯的影响  

Effect of high expression of PNPLA3 gene on cell proliferation and triglyceride in model of nonalcoholic fatty liver disease

作　　者：何晓萱 马雪儿 杨雪霞 李琴 贾茜茜 任树风 蔡雯[1] HE Xiaoxuan;MA Xueer;YANG Xuexia;LI Qin;JIA Qianqian;REN Shufeng;CAI Wen(School of Nursing,Xinjiang Medical University,Xinjiang Uygur Autonomous Region,Urumqi 830011,China)

机构地区：[1]新疆医科大学护理学院,新疆乌鲁木齐830011

出　　处：《中国医药导报》2024年第23期9-13,27,共6页China Medical Herald

基　　金：新疆维吾尔自治区2022年度省部共建中亚高发病成因与防治国家重点实验室代谢病专项项目(SKL-HIDCA-2022-DX5);新疆维吾尔自治区自然科学基金面上项目(2020D01C170)。

摘　　要：目的探讨patatin样磷脂酶域3(PNPLA3)高表达对非酒精性脂肪性肝病(NAFLD)细胞模型细胞增殖和甘油三酯(TG)的影响。方法将人肝癌细胞(SMMC-7721细胞)分为对照组、模型组、PNPLA3过表达组、PNPLA3过表达空载组。模型组:经游离脂肪酸诱导建立NAFLD细胞模型,PNPLA3过表达组、PNPLA3过表达空载组:在造模后经慢病毒原液感染、嘌呤霉素筛选获得。用定量反转录聚合酶链反应(qRT-PCR)、MTT比色法和TG测试技术检测细胞中PNPLA3表达情况、细胞存活率和胞内TG含量。结果qRT-PCR结果显示,模型组PNPLA3基因的表达量高于对照组(P<0.01),PNPLA3过表达组PNPLA3基因表达量高于模型组(P<0.01)。MTT结果显示,模型组细胞相对存活率高于对照组(P<0.05),PNPLA3过表达组细胞相对存活率高于模型组(P<0.01)。TG检测结果显示:模型组TG含量高于对照组(P<0.01),PNPLA3过表达组中胞内TG含量高于模型组(P<0.05)。结论成功构建PNPLA3过表达NAFLD稳转株,PNPLA3的过量表达有助于细胞脂质堆积并增强细胞相对存活率。Objective To investigate the effect of high expression of patatin-like phospholipase domain 3(PNPLA3)on proli-feration and triglyceride(TG)of nonalcoholic fatty liver disease(NAFLD)model cells.MethodsHuman hepatoma cells(SMMC-7721 cells)were divided into control group,model group,PNPLA3 overexpression group,and PNPLA3 overexpression noload group.Model group:NAFLD cell model was established by free fatty acid induction,and PNPLA3 overexpression group and PNPLA3 overexpress noload group obtained by lentivirus stock solution infection and puromycin screening after modeling.Quantitative reverse transcription polymerase chain reaction(qRT-PCR),MTT colorimetric assay,and TG testing were used to detect the expression of PNPLA3,cell viability,and intracellular TG content in cells.Results The results of qRT-PCR showed that the expression level of PNPLA3 gene in the model group was higher than that in the control group(P<0.01),and the expression level of PNPLA3 gene in the PNPLA3 overexpression group was higher than that in the model group(P<0.01).MTT results showed that the relative survival rate of cells in the model group was higher than that in the control group(P<0.05),and the relative survival rate of cells in the PNPLA3 overexpression group was higher than that in the model group(P<0.01).The results of TG detection showed that TG content in model group was higher than that in control group(P<0.01),the TG content in the PNPLA3 overexpression group was higher than that in the model group(P<0.05).Conclusion The stable NAFLD strain with PNPLA3 overexpression is successfully constructed,and the over-expression of PNPLA3 contributes to the accumulation of lipid and increases the relative survival rate of cells.

关 键 词：非酒精性脂肪性肝病 patatin样磷脂酶域3 基因表达 肝癌细胞 慢病毒 甘油三酯 

分 类 号：R34[医药卫生—基础医学]