【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。更多还原

【Abstract】 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.更多还原