大鼠骨髓间充质干细胞向肌成纤维母细胞的体内诱导  

In Vivo Induction of Differentiation of Rat Bone Marrow Mesenchymal Stem Cells into Myofibroblasts

作　　者：周伟[1] 徐艳华[2] 吴小翎[1] 姜蓉[2] 陈鹏飞[1] 

机构地区：[1]重庆医科大学附属第二医院消化内科,重庆400010 [2]重庆医科大学组织学与胚胎学教研室,重庆400016

出　　处：《中国生物制品学杂志》2011年第5期561-565,共5页Chinese Journal of Biologicals

摘　　要：目的探讨大鼠骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMSCs)在不同疾病模型中分化为肌成纤维母细胞的可能性。方法取大鼠股骨和胫骨,采用全骨髓差异贴壁法分离纯化BMSCs,并体外传代,取第3代BMSCs,经流式细胞仪鉴定细胞表面抗原,并对其进行成骨、成脂诱导。将35只SD大鼠分为5组:正常对照组(A组)、肝纤维化模型组(B组)、溃疡性结肠炎模型组(C组)、BMSCs肝纤维化移植组(D组)和BMSCs溃疡性结肠炎移植组(E组),每组7只。B组和D组大鼠经皮下注射40%四氯化碳复制肝纤维化模型,建模8周后,D组大鼠通过尾静脉移植经DAPI标记的大鼠BMSCs 1×106个;C组和E组大鼠采用三硝基苯磺酸(TNBS)-乙醇溶液灌肠,复制溃疡性结肠炎模型,灌肠后24 h,E组大鼠通过尾静脉移植DAPI标记的大鼠BMSCs 1×106个。于移植BMSCs 2周后处死所有大鼠,取B组和D组大鼠肝组织,经HE、VG染色观察肝脏病理变化;取C组和E组大鼠结肠组织,经HE染色观察肠组织病理变化;所有组织经免疫荧光法检测α-平滑肌肌动蛋白(α-smoothmuscle actin,α-SMA)的表达,激光共聚焦显微镜观察BMSCs在肝脏和肠道的分布,并观察α-SMA的分布。结果 BMSCs表达CD29、CD166和CD90,不表达CD45,并可进行成骨、成脂诱导。B组和D组大鼠肝组织建模10周后可见大鼠肝索排列紊乱,在门脉区有大量的胶原纤维增生,形成假小叶;C组和E组大鼠结肠可见黏膜增厚,形成溃疡,黏膜及黏膜下层可见大量炎性细胞浸润。DAPI标记的BMSCs主要分布在D组大鼠肝脏纤维条索中,并同时表达α-SMA;而E组大鼠结肠黏膜及黏膜下层中可见DAPI标记的BMSCs,这些细胞同时也表达α-SMA。结论 BMSCs在肝纤维化大鼠肝脏和溃疡性结肠炎大鼠结肠中均可分化为肌成纤维母细胞。Objective To investigate the possibilities of differentiation of rat bone marrow mesenchymal stem cells（BMSCs） into myofibroblasts in various disease models.Methods BMSCs were isolated and purified from the marrows in femora and tibiae of rats by whole bone marrow adherent culture method and subcultured in vitro.The BMSCs of passage 3 were identified for surface antigen and induced to evaluate the abilities of adipogenesis and osteogenesis.Thirty-five rats were divided into normal control（A）,liver fibrosis model（B）,ulcerative colitis model（C）,liver fibrosis model transplanted with BMSCs（D） and ulcerative colitis model transplanted with BMSCs（E） groups,seven for each.The rats in groups B and D were injected s.c.with 40% carbon tetrachloride to copy liver fibrosis model,and those in group D were transplanted with 1 × 106 DAPI-labeled BMSCs through caudal vein 8 weeks later.However,the rats in groups C and E were treated with ethanol solution containing trinitrobenzenesulfonate（TNBS） by enema to copy ulcerative colitis model,and those in group E were transplanted with 1 × 106 DAPI-labeled BMSCs through caudal vein 24 h later.All the rats were killed 2 weeks after transplantation with BMSCs,of which the liver tissues in groups B and D were observed for pathological change by HE and VG staining,while the intestinal tissues in groups C and E by HE staining.The expression of α-smooth muscle actin（α-SMA） in all the tissue samples were determined by IFA.The distribution of BMSCs in liver and intestinal tract and α-SMA in liver were observed by laser confocal microscopy.Results BMSCs were positive for CD29,CD166 and CD90 but negative for CD45,and were differentiated into osteoblasts and adipocytes.Ten weeks after copy of disease models,the hepatic cords of rats were arranged irregularly,with the proliferation of a large quantity of collage fibers in portal region and formation of pseudolobules.However,in groups C and E,the mucosal thickening of colon,ulcer and infiltration of a large qu

关 键 词：间质干细胞 肝硬化 结肠炎 溃疡性 细胞分化 成纤维细胞 

分 类 号：Q254[生物学—细胞生物学]