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当前位置: 首页 > 文章 > CTGF-siRNA对TGF-β1诱导的肝星状细胞胶原分泌的影响 中国食物与营养 2014,20 (08) 64-67
Position: Home > Articles > Effects of CTGF-siRNA on Expression of Extracellular Matrix in TGF-β1-induced Rat Hepatic Satellite Cells Food and Nutrition in China 2014,20 (08) 64-67

CTGF-siRNA对TGF-β1诱导的肝星状细胞胶原分泌的影响

作  者：
张羽飞;李厚忠;高晓虹;袁晓环;初彦辉
单  位：
牡丹江医学院生物技术实验室;牡丹江医学院药学院药理教研室;牡丹江医学院黑龙江省抗纤维化生物治疗重点实验室
关键词：
肝星状细胞;TGF-β1;结缔组织生长因子;siRNA;肝纤维化
摘  要：
目的:研究结缔组织生长因子(connective tissue growth factor,CTGF)的小RNA(CTGF-siRNA)对TGF-β1诱导的大鼠肝星状细胞(HSC-T6)胶原分泌的影响。方法:实验分为5组:正常组(单纯HSC-T6细胞组)、对照组(TGF-β1诱导组)、siRNA组、非特异siRNA组、lipo组(脂质体组)。培养大鼠HSC-T6细胞,以10ng/mL终浓度的TGF-β1对培养的HSC-T6细胞进行诱导,再通过脂质体法将CTGF-siRNA转染入HSC-T6细胞,通过RT-PCR、Western blotting方法研究各实验组细胞中CTGF、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、I型胶原蛋白(Collagen type I,Col 1a1)、纤维连接蛋白(fibronectin,FN)mRNA及蛋白的表达水平。结果:正常组细胞内CTGF有低水平的表达,在给予TGF-β1诱导后表达水平显著升高(P<0.05);TGF-β1刺激24h后,靶向转染入特异性siRNA后,CTGF、Col 1a1、FN及α-SMA的表达均显著下调(P<0.05)。结论:本研究提示,通过RNA干扰使CTGF基因沉默后,在体外可明显抑制HSC-T6肝成纤维细胞的表型转化及细胞外基质的合成,进一步提示CTGF是促肝纤维化的关键性细胞因子,而RNA干扰是一种特异高效的很有前途的干预手段,为基因治疗肝纤维化提供实验依据。
译  名：
Effects of CTGF-siRNA on Expression of Extracellular Matrix in TGF-β1-induced Rat Hepatic Satellite Cells
作  者：
ZHANG Yu-fei;LI Hou-zhong;GAO Xiao-hong;YUAN Xiao-huan;CHU Yan-hui;Heilongjiang Key Laboratory of Anti-fibrosis Biotherapy,Mudanjiang Medical University;Medical Research Center,Mudanjiang Medical University;Department of Pamacy,Mudanjiang Medical University;Department of Biotechnology,Mudanjiang Medical University;
关键词：
HSC-T6 cells;;TGF-β1;;Connective Tissue Growed Factor(CTGF);;siRNA;;liver fibrosis
摘  要：
【Objective】To study the effects of CTGF-siRNA on expression of extracellular matrix in TGF-β1-induced rat hepatic satellite cells( HSC). 【Method】Five groups were set as normal group( pure HSC-T6 control),model group( TGF-β1 induction group),siRNA group,scrambled-siRNA group,pure liposome supplement group. The HSC-T6 cells were cultured and stimulated with TGF-β1 at an end concentration of 10 ng /mL for 24 h,then the short hairpin RNA( siRNA) was transferred into the HSC-T6 cells. Lastly,the mRNA and protein levels of connective tissue growth factor( CTGF),α-smooth muscle actin( α-SMA),Collagen type I( Col 1a1) and fibronectin( FN) were detected by means of reverse transcriptional polymerse chain reaction( RT-PCR) and Western blotting. 【Result】Low level expressions of CTGF was detected in basal state. When stimulated by TGF-β1,the expression of CTGF were remarkably increased. After TGF-β1 treatment for 24 h,the expressions of CTGF,α-SMA,Col 1a1 and FN were inhibited prominently by siRNA that targeted to CTGF gene.【Conclusion】Our data suggested that chemically synthetic siRNA could silence CTGF and decline the deposition of extracellular matrix in CTGF-siRNA / HSC-T6 cells induced by TGF-β1,which suggested CTGF was an important cytokine in liver fibrosis and siRNA targeting CTGF gene might be a promising strategy for gene therapy in preventing liver fibrosis.

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