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不同途径移植骨髓间充质干细胞治疗肝硬化小鼠的效果比较
Bone marrow mesenchymal stem cell transplantation via different approaches in treatment of liver cirrhosis in mice
文章发布日期:2016年09月30日  来源:  作者:张丽霞,李莹,王黎明,等  点击次数:1532次  下载次数:263次

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【摘要】:目的评价经不同途径移植骨髓间充质干细胞(BMSC)治疗小鼠肝硬化的效果差异。方法46只BALB/c小鼠随机分为正常对照组5只和肝硬化模型组41只。用CCl4橄榄油皮下注射构建小鼠肝硬化模型,选取肝硬化小鼠36只,随机分为对照组、尾静脉移植组、脾内移植组,每组各12只。以全骨髓贴壁法获得第3代BMSC,通过流式细胞分析术行细胞表面鉴定,选取尾静脉及脾脏注射至肝硬化小鼠体内。移植后4周分别取血检测肝功能,取肝组织切片进行HE、Masson染色及α平滑肌肌动蛋白(αSMA)免疫组化检查,检测小鼠肝细胞组织损伤和纤维化情况。组间比较采用单因素方差分析。结果造模8周后,模型组小鼠毛发变稀,色泽暗黄,摄食减少,活动减少,体质量下降。细胞移植后,脾内移植和尾静脉移植组小鼠Alb较对照组明显升高[(2905±341) g/L vs (2103±218) g/L,(2946±325) g/L vs (2103±218) g/L],ALT、AST明显降低[(17067±3272) U/L vs (25458±4793) U/L,(16658±2824) U/L vs (25458±4193) U/L; (20217±3158) U/L vs (29308±3523) U/L, (18650±3313) U/L vs (29308±3523) U/L],差异均有统计学意义(P值均<0.01);两移植途径间比较差异均无统计学意义(P值均>0.05);组织病理改变显著,两移植组小鼠肝硬化程度减轻,胶原纤维及坏死面积减少,组织结构好,与对照组比较,肝脏胶原面积比值显著降低[(1436±221)% vs (26.38±082)%,(1088±127)% vs (2638±082)%](P值均<001)。免疫组化染色提示两移植组的αSMA阳性表达细胞数较对照组均明显下降(1440±169)% vs (256±073)%,(997±146)% vs (2560±073)%](P值均<001)。结论经两种途径移植BMSC对肝硬化小鼠肝功能、病理组织学等方面均有改善,二者效果相似。
【Abstract】:ObjectiveTo investigate the clinical effects of bone marrow mesenchymal stem cell (BMSC) transplantation via different approaches in the treatment of liver cirrhosis in mice. MethodsA total of 46 BALB/c mice were randomly divided into normal control group with 5 mice and liver cirrhosis model group with 41 mice. Subcutaneously injected carbon tetrachloride olive oil was used to establish the mouse model of liver cirrhosis. A total of 36 mice with liver cirrhosis were randomly divided into control group, caudal vein BMSC transplantation group, and spleen BMSC transplantation group, with 12 mice in each group. Whole bone marrow adherent culture was performed to obtain the third-generation BMSCs, and flow cytometry was used for cell surface identification. BMSCs were injected into the mice through the caudal vein or spleen. Blood samples were collected at 4 weeks after transplantation to measure liver function. HE and Masson staining and α-smooth muscle actin (α-SMA) immunohistochemistry were performed for liver sections. Liver injury and fibrosis in mice were examined. A one-way analysis of variance was used for comparison between groups. ResultsAt 8 weeks after the establishment of the model, the mice in the model group had sparse and dark yellow hair, reduced food consumption and activity, and a reduction in body weight. After transplantation, compared with the model control group, the caudal vein BMSC transplantation group and spleen BMSC transplantation group showed a significant increase in albumin and significant reductions in alanine aminotransferase and aspartate aminotransferase (all P<0.01). There were no significant differences between the two transplantation approaches (P>0.05). After transplantation, there were significant changes in diseased tissue, alleviated liver cirrhosis, reduced collagen fiber and necrotic area, and a good structure. Immunohistochemistry showed both transplantation groups showed significant reductions in the number of cells with positive α-SMA. ConclusionBMSC transplantation via the tail vein or spleen can improve liver function and diseased tissue in mice with liver cirrhosis and these two approaches have comparable clinical effects.
【关键字】:肝硬化; 间质干细胞移植; 小鼠, 近交BALB C
【Key words】:liver cirrhosis; mesenchymal, stem cells; transplantion; mice, inbred BALB C
【引证本文】:张丽霞, 李莹, 王黎明, 等. 不同途径移植骨髓间充质干细胞治疗肝硬化小鼠的效果比较[J]. 临床肝胆病杂志, 2016, 32(10): 1906-1910.

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