Objective:To observe the conditions of isolation,purification and culture of mesenchymal stem cells(MSCs) in vitro derived from human umbilical cord,and to observe the repairing effect on experimental hepatic fibrosis in rats by transplantation with MSCs only or MSCs with IL-10.Methods:The MSCs derived from human umbilical cord were isolated,purified and cultivated in vitro.The model of hepatic fibrosis induced by CCl4 was established.After the model of hepatic fibrosis was succeeded,the rats were divided into three groups:model group,MSCs transplantation group,MSCs transplantation with IL-10 group.MSCs transplantation group was given 1×106 MSCs by the tail vein injection at the first 1 weeks,2 weeks and 3 weeks.MSCs transplantation with IL-10 group not only was given MSCs the same with MSCs transplantation group, but also was injected of IL-10 ( 4 μg/kg ) in abdomen, 4 times a week.After 4 weeks,the rats were executed,rats blood was collected to observe the changes of liver function and the index of liver fibro-sis.The liver HE staining was carried out to observe the changes of liver pathology.Expression of MMP-2 mRNA was detected by RT-PCR method,expression of TGF-βprotein was detected by Western blot method.Results:The rat liver function and the index of liver fibrosis improved obviously with a significant difference between model group , MSCs transplantation group and MSCs transplantation with IL-10 group after transplanting,MSCs with IL-10 was better than MSCs only.Liver tissue of rats with hematoxylin eosin staining suggested,liver fibrosis was obviously improved,and the MSCs transplantation with IL-10 group was best.MMP-2 mRNA expression was significantly higher between normal group and model group,MSCs transplantation group with a significant difference (P<0.05),MMP-2 mRNA expression was higher with a no significant difference between model group and MSCs transplantation with IL-10 group ( P>0.05).TGF-βprotein expression of other groups was higher than normal group with a significant (P<0.05),and MSCs transplantation with IL-10 group was the largest decrease than normal group.Conclusion:MSCs only or MSCs with IL-10 transplantation can improve the biochemical characteristics of rat peripheral blood and liver histological structure,and with IL-10 wan better than MSCs only.MSCs and IL-10 play a role in the treatment of liver fibrosis through lowering the expression of MMP-2 and the expression of TGF-βprotein.%目的：探究人脐带源间充质干细胞（ MSCs）的分离、培养方法，通过对MSCs与IL－10联合治疗四氯化碳诱导的大鼠肝纤维化的研究，观察其协同效应，为临床治疗肝纤维化寻找新方法，并阐明其在肝纤维化治疗中的部分机制。方法：自然贴壁法分离、纯化人脐带间充质干细胞并进行体外培养和扩增。50只大鼠随机选取5只入选正常对照组（n＝5），其余45只大鼠采用皮下多点注射CCl4方法制备大鼠肝纤维化模型，造模成功后随机分为模型损伤组（n＝15）、细胞移植组（n＝15）和联合用药组（ n＝15）。细胞移植组在模型制备成功后的第1周、第2周、第3周经尾静脉给予1×106脐带间充质干细胞治疗；联合治疗组在给予细胞移植组相同的细胞治疗基础上，给予腹腔内注射IL－10（4μg／kg），每周4次。4周后将大鼠处死，收集各组大鼠血液检测肝功能、肝纤维化指标；摘取肝脏行苏木精－伊红染色，观察病理变化；RT－PCR法检测MMP－2 mRNA表达情况，Western blot法检测TGF－β蛋白的表达。结果：细胞移植组与联合治疗组大鼠肝功能、肝纤维化指标均明显改善，与对照组比较，差异有显著性意义（ P＜0．05），且联合治疗组改善更明显（ P＜0．05）；大鼠肝组织苏木精－伊红染色提示，细胞移植组、联合治疗组肝纤维化程度明显改善，联合治疗组肝脏病理更接近于正常；损伤组、细胞移植组MMP－2 mRNA表达明显增高，明显高于正常对照组（P＜0．05），联合治疗组MMP－2 mRNA表达增高，与正常对照组比较无显著性差异（P＞0．05）；与正常对照组比较，其他组TGF－β蛋白的表达都有所增高（P＜0．05），其中损伤组TGF－β蛋白的表达增高最明显（P＜0．05），细胞移植组、联合治疗组TGF－β蛋白的表达较损伤组均有不同程度的降低（P＜0．05），联合治疗组与细胞移植组比较TGF－β蛋白的表达也有显著降低（P＜0．05）。结论：脐带间充质干细胞单独移植或与IL－10联合治疗均可以改善肝纤维化大鼠外周血液的生化特性和肝的组织学结构，且IL－10联合治疗效果更佳，这可能是由于间充质干细胞与IL－10可下调MMP－2 mRNA的表达，保护肝小叶结构；抑制TGF－β蛋白的表达，减少肝星状细胞的激活等原因。
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