骨髓间充质干细胞移植治疗四氯化碳大鼠肝纤维化的实验研究  

Experimental study of bone marrow mesenchymal stem cells transplantation for treatment of liver fibrosis in rats with carbon tetrachloride

作　　者：郑雪 李静[1] 黄俊凯 丁新 陈卫刚[1] ZHENG Xue;LI Jing;Huang Junkai;DING Xin;CHEN Weigang(Department of Gastroenterology,First Affiliated Hospital of School ofMdedicine,Shihezi University,Shihezi,Xinjiang 832000,China)

机构地区：[1]石河子大学医学院第一附属医院消化内科

出　　处：《石河子大学学报（自然科学版）》2019年第5期636-641,共6页Journal of Shihezi University(Natural Science)

基　　金：新疆生产建设兵团社会发展与科技攻关计划项目(2015AD015)

摘　　要：目的通过移植大鼠骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMSC)治疗肝纤维化大鼠,并评价其疗效。方法通过全骨髓贴壁法分离培养纯化大鼠BMSC,用流式细胞仪对第3代BMSC进行表面抗原鉴定。选取清洁雄性SD大鼠20只,利用四氯化碳(Carbon tetrachloride,CCl 4)皮下注射构建大鼠肝纤维化模型,将肝纤维化大鼠随机分为BMSC组、PBS组,每组10只;分别通过尾静脉注射,予1 mL细胞浓度为1×10 6个/mL的BMSC细胞悬液、1mL PBS溶液,另取6只健康雄性SD大鼠作为空白对照组;移植后第5周处死各组大鼠,采集血液检测大鼠肝功;取肝脏组织切片进行HE、Masson染色,观测大鼠肝细胞组织损伤及纤维化程度,利用Image J软件计算肝脏胶原容积分数(Collagen Volume Fraction,CVF);通过qRT-PCR检测各组大鼠肝脏Ⅰ、Ⅲ型胶原mRNA表达。结果(1)通过流式细胞术鉴定BMSC表面抗原,CD29、CD90高表达,CD45低表达。(2)与PBS组比较,BMSC组大鼠肝脏肝小叶网状结构较完整,CVF降低且Ⅰ、Ⅲ型胶原mRNA表达减少(P<0.05)。(3)与PBS组相比,BMSC组大鼠肝功能改善,且差异具有统计学学意义(P<0.05)。结论同种异体移植BMSC可以有效减少肝脏胶原沉积,改善肝纤维化大鼠肝脏功能。Objective To treat liver fibrosis rats by transplanting rat bone marrow mesenchymal stem cells(BMSC)and evaluate their therapeutic effects.Methods Rat BMSCs were isolated and cultured by whole bone marrow adherence method.The surface antigens of the third generation BMSCs were identified by flow cytometry.Twenty male SD rats were selected and subcutaneously injected with carbon tetrachloride(CCl 4)to construct rat liver fibrosis model.Rats with hepatic fibrosis were randomly divided into BMSC group and PBS group,with 10 rats in each group.Through tail vein injection,1 mL of BMSC cell suspension with a concentration of 1×10^6/mL,1 mL of PBS solution,and 6 healthy male SD rats were used as blank control group;each week after transplantation,each group was sacrificed.Rats in the group were collected for blood test for liver function.Liver tissue sections were taken for HE and Masson staining.The degree of hepatocyte tissue damage and fibrosis was observed.The Collagen Volume Fraction(CVF)was calculated by Image J software.The expression of type I and III collagen mRNA in liver of each group was detected by qRT-PCR.Results(1)BMSC surface antigen was identified by flow cytometry,CD29 and CD90 were highly expressed,and CD45 was lowly expressed.(2)Compared with PBS group,the hepatic lobular reticular structure of BMSC group was intact,CVF decreased and typeⅠandⅢcollagen mRNA The expression was decreased(P<0.05).(3)Compared with the PBS group,the liver function of the BMSC group was improved,and the difference was statistically significant(P<0.05).Conclusion Allogeneic transplantation of BMSC can effectively reduce liver collagen deposition and improve liver function in rats with liver fibrosis.

关 键 词：骨髓间充质干细胞 大鼠肝纤维化 细胞移植 

分 类 号：R678.73[医药卫生—外科学]