人羊膜脐带源性间充质干细胞对大鼠肝硬化的治疗作用

Therapeutic effect of transplantation of human amniotic membrane-and umbilical cord-derived mesenchymal stem cells on hepatic cirrhosis in rats

目的:比较人脐带间充质干细胞(human amniotic membrane-derived mesenchymal stem cells,hAM-MSCs)和人羊膜间充质干细胞(human umbilical cord-derived mesenchymal stem cells,hUC-MSCs)对模型大鼠肝硬化的治疗效果.方法:分离培养hAM-MSCs和hUC-MSCs,流式检测CD29、CD44和CD34.CCl4诱导大鼠肝硬化模型,在第8周时,杀死5只大鼠做病理检测以确诊为肝硬化.30只成模大鼠随机分为脐带干细胞组(n=10)、羊膜干细胞组(n=10)和对照组(n=10),分别注入hAM-MSCs、hUC-MSCs2mL(细胞数量2×106)和等量生理盐水.细胞移植前和移植4wk后,检测大鼠肝功能;肝组织HE染色、Masson染色;免疫组织化学法检测α-SMA在肝脏中的表达.结果:2种细胞表达CD29、CD44,不表达CD34.移植后,与对照组相比,脐带干细胞组和羊膜干细胞组ALT和AST明显降低(204.6±16.4,195.6±21.2vs539.8±36.2;180.1±25.2,167.5±19.0vs337.4±23.4,P<0.05);胶原沉积减少;组织病理学评分有显著性差异(P<0.05);肝脏α-SMA表达量减少(130.6±3.0,127.0±2.6vs152.2±5.4,P<0.05).脐带干细胞组和羊膜干细胞组的肝功能、肝组织病理学评分及肝脏α-SMA表达量均无明显差异.结论:hAM-MSCs和hUC-MSCs能有效改善肝硬化大鼠模型的肝功能和肝硬化程度,两者治疗效果无明显差异.

AIM：To evaluate the therapeutic effect of trans-plantation of human amniotic membrane-derived mesenchymal stem cells（hAM-MSCs） and human umbilical cord-derived mesenchymal stem cells（hUC-MSCs） on carbon tetrachloride（CCl4）-induced hepatic cirrhosis in rats.METHODS：hAM-MSCs and hUC-MSCs were isolated and analyzed by flow cytometry for detection of expression of CD44,CD29 and CD34.Hepatic cirrhosis was induced in rats with CCl4.At week 8,five rats were killed to conduct pathological examination to confirm successful induction of hepatic cirrhosis,and 30 rats with hepatic cirrhosis were randomly and equally divided into three groups：hAM-MSCs group,hUC-MSCs group and control group.The hAMMSCs and hUC-MSCs groups were infused wit 2×106 MSCs in 2 mL of saline via the tail vein,while the control group was given equal volume of saline.Liver function was examined before cell transplantation and 4 wk after cell transplantation.HE staining and Masson dyeing were performed to observe pathological changes in the liver.The expression of alpha-smooth muscle actin（α-SMA） in the liver was determined by immunohistochemistry.RESULTS：Both isolated hAM-MSCs and hUCMSCs expressed CD29 and CD44,but did not express CD34.After cell transplantation,liver function parameters were markedly improved（all P 0.05） and the expression of α-SMA was reduced in the hAM-MSCs and hUC-MSCs groups compared to the control group（130.6 ± 3.0,127.0 ± 2.6 vs 152.2 ± 5.4,both P 0.05）.There were no statistically significant differences in liver function parameters and α-SMA expression between the hAM-MSCs and hUC-MSCs groups.CONCLUSION：Transplantation of hAM-MSCs and hUC-MSCs could efficiently improve liver functions and inhibit liver fibrosis in rats.