中国医药导报2024,Vol.21Issue(23) :9-13,27.DOI:10.20047/j.issn1673-7210.2024.23.02

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

何晓萱 马雪儿 杨雪霞 李琴 贾茜茜 任树风 蔡雯
中国医药导报2024,Vol.21Issue(23) :9-13,27.DOI:10.20047/j.issn1673-7210.2024.23.02

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

何晓萱 1马雪儿 1杨雪霞 1李琴 1贾茜茜 1任树风 1蔡雯1
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作者信息

  • 1. 新疆医科大学护理学院,新疆乌鲁木齐 830011
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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.

Abstract

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

Key words

Nonalcoholic fatty liver disease/Patatin-like phos-pholipase domain 3/Gene expression/Hepatoma carcinoma cells/Lentivirus/Triglyceride

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出版年

2024
中国医药导报
中国医学科学院

中国医药导报

CSTPCD
影响因子:1.759
ISSN:1673-7210
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