PNPLA3 I148M基因突变对大鼠肝星状细胞转化生长因子β1表达的影响

目的探讨patatin样磷脂酶域（PNPLA3）I148M基因突变在非酒精性脂肪性肝纤维化发生发展中的作用机制。方法构建携带PNPLA3 I148M基因突变型和野生型的慢病毒载体,转染大鼠肝星状细胞（HSC-T6）,采用实时荧光定量PCR的方法检测转化生长因子（TGF）β1 mRNA的表达水平。采用t检验进行组间比较。结果构建了携带PNPLA3 I148M突变型和野生型的慢性病毒载体,并成功转染HSC-T6细胞,建立了能够稳定表达PNPLA3突变型和野生型基因的HSC-T6细胞系;PNPLA3 I148M突变型与野生型相比,TGFβ1 mRNA相对表达量明显上调,差异具有统计学意义（1.25±...

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Published in临床肝胆病杂志 Vol. 32; no. 4; pp. 769 - 771
Main Author 安佰全 辛永宁 芦琳琳 宣世英
Format Journal Article
LanguageChinese
Published 青岛市市立医院消化内二科,山东青岛266021 2016
南京医科大学青岛临床医学院,山东青岛,266021%南京医科大学青岛临床医学院,山东青岛266021
青岛市消化疾病重点实验室,山东青岛266021%青岛市消化疾病重点实验室,山东青岛,266021
Subjects
点突变
磷脂酶类
肝硬化
转化生长因子β1
磷脂酶类
phospholipases
肝硬化
点突变
transforming growth factor betal
liver cirrhosis
point mutation
转化生长因子β1
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ISSN1001-5256
DOI10.3969/j.issn.1001-5256.2016.04.035

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Summary:目的探讨patatin样磷脂酶域（PNPLA3）I148M基因突变在非酒精性脂肪性肝纤维化发生发展中的作用机制。方法构建携带PNPLA3 I148M基因突变型和野生型的慢病毒载体,转染大鼠肝星状细胞（HSC-T6）,采用实时荧光定量PCR的方法检测转化生长因子（TGF）β1 mRNA的表达水平。采用t检验进行组间比较。结果构建了携带PNPLA3 I148M突变型和野生型的慢性病毒载体,并成功转染HSC-T6细胞,建立了能够稳定表达PNPLA3突变型和野生型基因的HSC-T6细胞系;PNPLA3 I148M突变型与野生型相比,TGFβ1 mRNA相对表达量明显上调,差异具有统计学意义（1.25±0.15 vs 0.48±0.07,t=11.826,P〈0.001）。结论PNPLA3 I148M基因突变能够促进HSC中TGFβ1的表达,为进一步探讨PNPLA3 I148M基因突变在非酒精性脂肪性肝纤维化中的作用提供了细胞模型和新的研究思路。
Bibliography:liver cirrhosis; transforming growth factor beta1; phospholipases; point mutation
AN Baiquan, XIN Yongning, LU Linlin （ Qingdao Medical College of Nanjing Medical University, Qingdao, Shandong 266021, China）
Objective To investigate the mechanism of action of PNPLA3 I148 M mutation in the development and progression of non- alcoholic fatty liver fibrosis. Methods The lentiviral vectors carrying the mutant or wild- type PNPLA3 I148 M gene were constructed and transfected into rat hepatic stellate（ HSC- T6） cells. Quantitative real- time PCR was applied to measure the mRNA expression of transforming growth factor β1（ TGF β1）. The t- test was applied for statistical analysis. Results The lentiviral vectors carrying the mutant or wild- type PNPLA3 I148 M gene were successfully constructed and transfected into HSC- T6 cells,and a HSC- T6 cell line with stable expression of the mutant or wild- type PNPLA3 gene was established. Compared with the cell line carrying the wild- type gene,the cell line carrying the mutant
ISSN:1001-5256
DOI:10.3969/j.issn.1001-5256.2016.04.035