CAJ | 학술논문

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.
목적 탐토IL-10(백개소-10)기인치료시부구유항소서간섬유화적작용급기궤제.방법 우음수중첨가류대을선알이유발소서간섬유화형성,수후경유전천공기인전송법장인류IL-10표체재체도입,채용면역조화법검측IL-10재소서간장중적표체;채용Sircol효원단백측정법검측간장중효원단백적함량;채용RT-PCR검측간섬유화상관기인[전화생장인자β1(TGF-β1)、효원단백α1(collagen α1)、종류배사인자-α(TNF-α)、세포간점련분자-1(ICAM-1)、섬련단백(Fibronectin)、혈관세포간점련분자-1(VCAM-1)、기질금속단백매억제인자(TIMP-1、TIMP-2)]적표체.결과 면역조화결과현시,IL-10기인치료가강저간섬유화정도.Sircol효원단백측정법현시,IL-10기인치료가감소효원단백적함량(P＜0.05).반전록취합매련반응현시IL-10기인치료가강저TGF-β1、TNF-α、ICAM-1화TIMPs mRNA적표체.결론 전천공IL-10기인치료가제공간경화일개신영적치료도경,병구유림상응용적잠력.
Objective 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.