摘要目的 探讨IL-10(白介素-10)基因治疗是否具有抗小鼠肝纤维化的作用及其机制.方法 于饮水中添加硫代乙酰胺以诱发小鼠肝纤维化形成,随后经由电穿孔基因传送法将人类IL-10表达载体导入,采用免疫组化法检测IL-10在小鼠肝脏中的表达;采用Sircol胶原蛋白测定法检测肝脏中胶原蛋白的含量;采用RT-PCR检测肝纤维化相关基因[转化生长因子β1(TGF-β1)、胶原蛋白α1(collagen α1)、肿瘤坏死因子-α(TNF-α)、细胞间黏连分子-1(ICAM-1)、纤连蛋白(Fibronectin)、血管细胞间黏连分子-1(VCAM-1)、基质金属蛋白酶抑制因子(TIMP-1、TIMP-2)]的表达.结果 免疫组化结果显示,IL-10基因治疗可降低肝纤维化程度.Sircol胶原蛋白测定法显示,IL-10基因治疗可减少胶原蛋白的含量(P＜0.05).反转录聚合酶链反应显示IL-10基因治疗可降低TGF-β1、TNF-α、ICAM-1和TIMPs mRNA的表达.结论 电穿孔IL-10基因治疗可提供肝硬化一个新颖的治疗途径,并具有临床应用的潜力.

abstractsObjective To investigate whether the interleukin-10 (IL-10) gene therapy has the effect of anti liver fibrosis in mice and its mechanism.Methods Liver fibrosis was induced by long-term thioacetamide administration in mice.Human IL-10 expression plasmid was delivered via electroporation after liver fibrosis was established.The immunohistochemistry was used to study the expression of IL-10 in liver.Sircol collagen determination method was used to detect the contents of collagen in the liver.The reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expressions of liver fibrosis-related genes,including transforming growth factor-β1 (TGF-β1),collagen α1,tumor necrosis factor-α (TNF-α),intercellular adhesion molecule-1 (ICAM-1),fibronectin,vascular cell adhesion molecule 1 (VCAM-1),and matrix metalloproteinase-inhibiting factor (TIMP-1,TIMP-2).Results Immunohistochemical results showed IL-10 gene therapy reversed hepatic fibrosis.Sircol collagen assay showed that IL-10 gene therapy reduced the content of collagen fibers(P ＜ 0.05).RT-PCR revealed IL-10 gene therapy reduced liver TGF-β1,TNF-α,collagen α1,cell adhesion molecule,and TIMPs mRNA upregulation.Conclusions Electroporative IL-10 gene therapy might be an effective therapeutic reagent for liver fibrosis with potential future clinical applications.