反义抑制结缔组织生长因子对实验性肝纤维化的影响

目的观察结缔组织生长因子(CTGF)反义RNA对实验性肝纤维化的影响。方法应用分子生物学技术构建CTGF反义RNA真核表达质粒,经腹腔注射入CCl4诱导的肝纤维化模型大鼠体内;通过RT-PCR、Western blot及免疫组织化学等方法检测CTGF反义RNA基因转染前后CTGF mRNA和蛋白质在肝组织中含量的变化;并通过检测肝组织中Ⅰ、Ⅲ型胶原含量的变化,观察CTGF反义RNA对大鼠肝纤维化的影响。结果转染CTGF反义RNA后,可抑制CTGF mRNA和蛋白质的表达(P<0.01),减少Ⅰ、Ⅲ型胶原的沉积(P<0.01),并在一定程度上促进肝组织的改善(P<0.01)。结论CTGF反义RNA对肝纤维化有一定的治疗作用。

Effects of antisense RNA of connective tissue growth factor expressing plasmid on rat liver fibrosis

Objective To observe the effects of antisense RNA of connective tissue growth factor (CTGF) on rat liver fibrosis. Methods Gene recombinant techniques were used to construct a rat antisense RNA of CTGF recombinant plasmid which could be expressed in eukaryotic cells. The recombinant plasmids were encapsulated with lipofectamin and then transducted into a carbon tetrachloride (CCU) induced rat liver fibrosis model. Expression of CTGF was assessed by RT-PCR, Western blot and immunohistochemistry. Immunohistochemistry was used to identify type I and III collagens. HE stained liver slides were used for pathological study. Results The mRNA and protein expression of CTGF in the fibrotic liver transfected with antisense-CTGF were significantly decreased compared with those of the controls (P < 0.01). The depositions of type I and type III collagens were also decreased (P < 0.05). Antisense-CTGF also minimized the pathological fibrosis in the rat livers (P < 0.01). Conclusion The results demonstrate that the antisense RNA of CTGF recombinant plasmid has certain effects in preventing liver fibrosis and makes it a possible candidate for use in future gene therapy.