腺病毒载体RNA干扰抑制结缔组织生长因子表达及其抗纤维化作用  

Inhibition of CTGF expression via adenovirus vector of RNA interference and its role against liver anti-fibrosis in vitro

作　　者：郝春秋[1] 彭梅娟[1] 谢玉梅[1] 周云[1] 魏欣[1] 马力[1] 王九萍[1] 张岩[1] 白雪帆[1] 贾战生[1] 

机构地区：[1]第四军医大学唐都医院感染病诊疗中心,西安710038

出　　处：《肝脏》2012年第12期854-857,共4页Chinese Hepatology

基　　金：国家自然科学基金资助项目(30571658)

摘　　要：目的探讨结缔组织生长因子(CTGF)基因沉默的抗纤维化作用。方法以介导CTGF基因沉默的腺病毒载体Ad.H1-CTGF感染HSC-T6细胞系,利用Real-time PCR及Western印迹在mRNA及蛋白质水平上观察CTGF基因沉默前后,CTGF自身及肝纤维化相关指标金属蛋白酶组织抑制剂-1(TIMP-1)、TIMP-2及I型胶原(Col-Ⅰ)表达水平的变化。结果腺病毒载体Ad.H1-CTGF感染HSC-T6后,CTGFmRNA转录及蛋白表达分别下调85.32%和76.07%;肝纤维化相关指标TIMP-1、TIMP-2及COL-Ⅰ在mRNA转录(下调82.34%、56.93%和74.21%)及蛋白质表达(下调76.53%、64.02%和70.79%)水平亦显著下调。结论腺病毒载体Ad.H1-CTGF可有效抑制HSC-T6中CTGF的表达,导致肝纤维化相关指标TIMP-1、TIMP-2及COL-Ⅰ的表达明显下调,可能具有潜在的抗纤维化作用。Objective To investigate the effects of knockdown of CTGF by adenovirus- delivered siRNA and anti- fibrosis in vitro. Methods CTGF was knocked down by adenovirus delivered siRNA in HSC-T6. The transcription of mRNA and expression of protein was detected with real-time PCR and western blot about CTGF itself and other fibrosis related indicators such as TIMP-1 （tissue inhibitor of metalloproteinase-1）, TIMP-2 and Col- I （collagen type I）. Results After HSC-T6 cells infected by adenovirus vector （Ad. H1-CTGF）, the CTGF had a suppressed mRNA and protein expression （85.32 %, 76.07 % ）. The transcription of mRNA and expressions of protein in TIMP-1 （82.34%, 76. 53 %）, TIMP-2（56.93%, 64.02 %） and Col-I （74. 21%, 70. 79 %） were significantly restrained by the inhibition of CTGF. Conclusion The adenovirus vector Ad. H1-CTGF has a better gene silencing effect targeting CTGF in HSC-T6. Gene silencing can further inhibit the TIMP-1, TIMP-2 and Col-I gene expression in HSC-T6. It has a positive significance in anti- fibrosis studying.

关 键 词：结缔组织生长因子 RNA干扰 腺病毒载体 抗纤维化 

分 类 号：R575.2[医药卫生—消化系统]