摘要
目的探讨骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMSCs)移植后对实验性肝纤维化大鼠的作用及其机制。方法直接贴壁法分离纯化雄性SD大鼠BMSCs;将30只雌性SD大鼠分为3组:正常组、肝纤维化模型组和BMSCs移植组,模型组和BMSCs移植组经皮下注射40%四氯化碳(CCL4),每周3次,复制大鼠肝纤维化模型,建模8周后,BMSCs移植组经尾静脉移植2×106个BMSCs,3 d后再次移植相同剂量的BMSCs,12周后处死各组大鼠,取外周血,分离血清,检测谷丙转氨酶(ALT)、谷草转氨酶(AST)和白蛋白(ALB)水平;取肝组织进行HE、VG染色,观察肝脏病理变化;免疫组化法和荧光定量PCR法检测肝组织中Ⅰ型胶原(TypeⅠcollagen,ColⅠ)和胶质纤维酸性蛋白(Glial fibrillary acidic protein,GFAP)的表达,荧光定量PCR法检测TGFβ1和Smad3 mRNA的表达。结果 BMSCs移植组较模型组大鼠肝组织纤维化程度明显加重;BMSCs移植组ALT和AST水平较模型组明显升高(P<0.05),ALB水平较模型组明显降低(P<0.05);ColⅠ和GFAP在模型组和BMSCs移植组纤维条索中有大量表达,BMSCs移植组中ColⅠ和GFAP mRNA及蛋白的表达量明显高于模型组(P<0.05);模型组和BMSCs移植组中TGFβ1和Smad3 mRNA的表达量明显高于正常组(P<0.05),且BMSCs移植组中TGFβ1和Smad3mRNA的表达量明显高于模型组(P<0.05)。结论 BMSCs移植可加重大鼠肝纤维化,BMSCs通过上调TGFβ/Smad信号传导通路的TGFβ1和Smad3的表达促进肝纤维化的进展。
Objective To investigate the effect of bone marrow mesenchymal stem cells （BMSCs） transplanted on experimental liver fibrosis in rats as well as the relevant mechanism. Methods BMSCs were isolated and purified from male SD rats by direct adherence method. Thirty female SD rats were divided into normal control, liver model and BMSC groups. The rats in model and BMSC groups were injected s. c. with 40% carbon tetrachloride, 3 times a week, to copy liver fibrosis model. Eight weeks after starting copy of the model, the rats in BMSC group were transplanted twice with 2 x 106 BMSCs, at an interval of 3 d. The rats in various groups were killed 12 after starting copy of the model, of which the alanine amiotransferase （ALT）, asparatate aminotransferase （AST） and albumin （ALB） content in sera were determined, the pathological change of liver was observed by HE and VG staining, the expressions of type I collagen （Col I ） and glial fibrillary acidic protein （GFAP） in liver were determined by immunohistochemical assay and fluorescent quantitative PCR, and those of transformation growth factor （TGF）β1 and Smad3 mRNAs by fluorescent quantitative PCR. Results The liver fibrosis of rats was severer in BMSC group than in model group. Compared with those in model group, the ALT and AST levels in BMSC group increased significantly （P 〈 0. 05）, while ALB level decreased significantly （P 〈 O. 05）. Col I and GFAP were expressed in a large quantity in fibers of rats in BMSC group, of which the mRNA and protein levels were significantly higher in BMSC group than in model group （P 〈 0. 05）. The expression levels of TGFβ1 and Smad3 mRNAs were significantly higher in model and BMSC groups than in normal group （P 〈 0. 05）, and in BMSC group than in model group （P 〈 0. 05）. Conclusion The transplantation with BMSCs aggravated the liver fibrosis of rats by up-regulating the expressions of TGFβ1 and Smad3 in TGFβ/Smad signal transduction pathway.
出处
《中国生物制品学杂志》
CAS
CSCD
2012年第2期176-180,共5页
Chinese Journal of Biologicals
关键词
间质干细胞移植
肝硬化
转化生长因子Β1
Smad3蛋白质
Mesenchymal stem cell transplantation
Liver cirrhosis
Transformation growth factor 131
Smad3 protein
