脂肪特异性蛋白27基因抑制大鼠肝星状细胞增殖活化  

Fsp27 gene inhibits the proliferation and activation of hepatic stellate cells in vitro

作　　者：俞富祥[1] 宋才鑫[1] 吴志伟[1] 朱千东[1] 张启瑜[1] 

机构地区：[1]浙江省温州医学院附属第一医院肝胆胰外科,325000

出　　处：《中华肝胆外科杂志》2013年第9期701-705,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：温州市科技计划项目（Y20120191）、浙江省重中之重外科学组资助（浙教高科2008-255）

摘　　要：目的探讨脂肪特异性蛋白27（Fsp27）基因对活化态肝星状细胞（HSCs）增殖、活化的影响及其对纤维化相关基因的调节作用。方法提取HSCs并培养。用实时定量PCR技术检测原代HSCs及活化HSCs中的Fsp27基因表达。构建携带Fsp27目的基因的慢病毒，转染活化的HSCs并继续培养72h。通过CCK-8比色法检测Fsp27基因对HSCs增殖的影响。Western blot检测HSCs α-肌动蛋白（α—SMA）的表达，了解HSCs的活化状态。实时定量PCR技术研究Fsp27基因对HSCs纤维化相关基因表达的影响。结果成功分离原代HSCs，Fsp27基因在原代HSCs及活化HSCs中表达差异显著（P〈0．01）。活化HSCs成功转染携带Fsp27目的基因的慢病毒后继续培养72h，与对照组比较，HSCs的活化与增殖受到明显抑制（P〈0．05）；Fsp27基因促进MMP-2的表达（P〈0．05），降低了TIMP-1及TGF-β1的表达（P〈0．05）。结论Fsp27基因具有抑制HSCs活化增殖的潜能，Fsp27基因可调节纤维化相关基因的表达。Fsp27基因的作用可能与维持HSCs静息状态细胞表型有关。Objective To investigate the Fsp27 gene＇s influence on the regulation of hepatic stellate cells （HSCs） in vitro. Methods HSCs were isolated from rat liver, the Fsp27 gene was detected in primary HSCs, and activated HSCs were detected by RT-qPCR. After 72 h of Fsp27 transduction through a lentivirus expressing Fsp27 （pLV-Fsp27）, the proliferation of HSCs was tested by the CCK-8 test kit, smooth muscle α-actin （α-SMA） expression of HSCs was tested by Western blot, and the fibrosis-related genes were tested by RT qPCR. Results The HSCs were isolated and cultured successfully, and the Fsp27 genetic difference between primary and activated HSCs was significant （P〈0.01）. After coculture for 72 h, Fsp27 inhibited the proliferation and activation of HSCs （P〈0.05）. Fsp27 can enhance expression of the MMP-2 gene and down-regulate expression of the TIMP-1 and TGF-β1 gene in activated HSCs （P〈0.05）. Conclusion The Fsp27 gene can inhibit the proliferation and activation of HSCs, regulate the expression of fibrosis-related genes, and may play an important role in maintaining the quiescent phenotype of HSCs.

关 键 词：脂肪特异性蛋白27 基因 肝星状细胞 增殖 活化 

分 类 号：R575.2[医药卫生—消化系统]