骨髓间充质干细胞可溶性成分对大鼠肝再生及血管内皮生长因子表达的影响  

Effects of soluble components derived from bone marrow mesenchymal stem cells on the expression of vascular endothelial growth factor and liver regeneration in rats

作　　者：丁爱兴[1] 禹亚彬[1] 黄庆峰[1] 吴宁[1] 卞建民[1] 

机构地区：[1]南京医科大学附属南京医院普外科,210006

出　　处：《中华肝胆外科杂志》2013年第10期762-766,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：南京市科技发展计划项目（200301094）;南京市医学科技发展专项资金资助项目（YKK10090）

摘　　要：目的观察骨髓间充质干细胞可溶性成分（BMSCs）对70％门静脉分支结扎诱导的大鼠血管内皮生长因子（VEGF）的表达及肝再生的影响。方法超声裂解提取BMSCs，经脾内注射至大鼠模型，同时设PBS对照组。我们对肝脏再生指数、肝脏细胞增殖、肝功能、再生肝脏的组织病理及肝性基因的表达进行分析，同时采用Western印迹检测及免疫组化检测VEGF的表达。结果术后2d和5d，BMSCs组的肝脏再生指数明显高于对照组（P〈0．05）；肝细胞的增殖细胞核抗原表达量是对照组的2倍左右（P〈0．05）；血清丙氨酸转氨酶及天冬氨酸转氨酶水平与对照组接近（P〉0．05）。组织病理表明，肝窦扩张及肝细胞空泡化比对照组轻；肝细胞增殖及血管生成相关基因的表达上调（P〈0．05）。BMscs组VEGF蛋白的相对表达量与对照组比较，差异无统计学意义（P〉0．05）。术后2d，BMSCs组门静脉周围和中心静脉周围的肝细胞均可见VEGF阳性表达，而对照组仅门静脉周围的肝细胞可见VEGF表达。结论BMSCs能够促进肝细胞增殖及VEGF介导的血管生成，维持肝细胞及肝脏结构的稳定，促进大鼠早期肝再生，为肝再生研究提供新途径。Objective To investigate the effects of soluble components derived from bone mar- row mesenchymal stem cells （BMSCs） on the expression of vascular endothelial growth factor （VEGF） and liver regeneration caused by 70% portal branch ligation （PBL） in rats. Methods Isolated and cultured BMSCs were lysed by sonication. PBL was performed in male SD rats followed by splenic injection of BMSCs or PBS as control. Animals were analyzed for liver regeneration index, hepatocytes proliferation, hepatic function, histopathological changes, and hepatic genes expression. Expression of VEGF was assessed by Western blot and immunohistochemistry. Results The liver regeneration index increased in the BMSCs group especially 2 and 5 days after PBL compared with the control group （P〈0.05） and reached （51.71±1.62）% and （76.82±0.81）% respectively. A 2-fold increase was showed in the PCNA labeling index of hepatocytes in rats treated with BMSCs compared with the control group （P〈0.05）. Histopathological findings showed that vacuolar change and sinusoidal congestion were lower in the BMSCs group. Alanine transaminase （ALT） and Aspartate transferase （AST） showed no significant difference between the two groups （P〈0.05）. On post operation day 2, hepatic interleukin-6 （IL6）, tumor necrosis factor α （TNFα）, hepatocyte growth factor （HGF）, vascular endothelial growth factor A （VEGFA）, and vascular endothelial growth factor 2 （VEGFR2） mRNAs tended to increase in the BMSCs group （P^0.05） while transforming growth factor β1 （TGF-β1） mRNA decreased （P〈0.05）. Western blot showed that the expression level of VEGF in the two groups were equal 2 and 5 days after surgery （P〉0.05）. On day 2 post operation, positive VEGF immunoreactivity was present in both pericentral and periportal hepatocytes in the BMSCs group, while only in periportal hepatocytes in the control group. Conclusion These results demonstrate that BMSCs accelerated liver regeneration caused by PBL

关 键 词：肝细胞 间充质干细胞 移植 肝功能试验 血管内皮生长因子 

分 类 号：R657.3[医药卫生—外科学]