脐带间充质细胞输注治疗大鼠肝硬化模型后表达谱的变化

Gene expression profile changes induced upon umbilical cord mesenchymal cell infusion therapy in a rat model of hepatic cirrhosis

目的 研究采用人脐带间充质细胞（UC-MSC）治疗肝硬化大鼠后,肝细胞中基因表达的变化. 方法 首先分离培养UC-MSC,然后鉴定其细胞表型以及向成骨细胞和脂肪细胞的分化能力.其次利用皮下注射四氯化碳方法建立大鼠肝硬化动物模型,将U C-MSC通过尾静脉注射途径对肝硬化大鼠进行治疗,利用血清学检测和组织学染色评价其治疗效果,最后提取肝脏RNA,进行表达谱芯片杂交并对结果进行统计分析,组间比较用t检验. 结果 细胞表型检测和诱导分化实验证明培养得到的贴壁细胞表型符合间充质细胞特征.血清学检测和组织病理学染色结果表明利用四氯化碳法成功建立了大鼠肝硬化模型,且U C-MSC治疗可以显著地改善肝损伤.芯片分析结果显示与对照组相比,脐带间充质细胞上调了补体凝血相关基因,下调细胞增殖、细胞周期和胶原合成等相关基因;肝功能生物化学指标检测显示球蛋白改善明显,分别为（33.9±1.5） g/L比（27.7±0.6） g/L,P=0.045;其他指标变化不明显. 结论 在脐带MSC治疗肝硬化逆转肝损伤的过程中,可能通过上调了补体凝血相关基因,抑制细胞增殖和胶原沉着发挥治疗作用.

Objective To investigate changes in gene expression that occur upon treatment with human umbilical cord mesenchymal stem cells （UC-MSCs） for hepatic cirrhosis using a rat model system.Methods Hepatic cirrhosis was induced in Sprague-Dawley rats by subcutaneous injection of carbon tetrachloride and oral administration of alcohol.UC-MSCs were isolated from human umbilical cord and the cells＇ immunophenotype and differentiation towards osteogenic and adipogenic lineages were confirmed.The UC-MSC sample or vehicle alone （phosphate buffered saline,PBS） was transplanted by intravenous injection.Histopathological staining and serological testing were used to compare the liver morphology and function among the different groups.The gene expression in the PBS group and UC-MSC group were detected by gene microarray and differences between the groups were statistically analyzed by t-test.Results Transplantation of the UC-MSCs improved liver function in the hepatic cirrhosis rats.Comparison of the gene expression profiles of the PBS group and the UC-MSC group showed that the latter had up-regulation of the genes related to the complement and coagulation cascades and down-regulation of the genes related to cell proliferation,cell cycle,and collagen synthesis.Conclusion UC-MSC therapy might improve liver function in cirrhosis by increasing the expression of genes related to the complement and coagulation cascades and by decreasing genes involved in cell proliferation and collagen deposition.