腺病毒介导的HIF-1α基因对人骨髓间充质干细胞分化成心肌细胞的影响  

Effects of Adenovirus- mediated Mutants of Hypoxia- inducible Factor 1α on Differentiation of Human Mesenchymal Stem Cells into Cardiomyocytes

作　　者：刘城[1] 吴平生[1] 王月刚[1] 裴静娴[1] 赖艳娴[1] 

机构地区：[1]广州市第一人民医院心血管内科,广东广州510180

出　　处：《湖北民族学院学报（医学版）》2013年第3期1-4,8,共5页Journal of Hubei Minzu University(Medical Edition)

基　　金：广东省自然科学基金(S2011040004458);广州市医药卫生科技项目(20121A011006)

摘　　要：目的研究腺病毒介导的野生型及突变型低氧诱导因子1α(hypoxia inducible factor-1α,HIF-1α)基因对体外诱导环境人骨髓间充质干细胞(human mesenchymal stem cells,hMSCs)分化成心肌细胞(cardiomyocyte)中的作用。方法重组腺病毒Ad-LacZ、Ad-HIF-1αnative、Ad-HIF-1α564、Ad-HIF-1α564/402及Ad-HIF-1α564/803分别以最佳转染复数转染体外5-氮胞苷(5-azacytidine,5-aza)诱导的hMSCs,于诱导培养后第6周提取细胞浆蛋白,应用酶联免疫分析检测心肌肌钙蛋白I(cTnI)含量,Western blot测定HIF-1α、VEGF及Nkx2.5蛋白表达水平。结果经5-aza诱导hMSCs第6周cTnI值为:对照组(0.700±0.013)、Ad-LacZ组(0.702±0.006)、Ad-HIF-1αnative组(0.729±0.004)、Ad-HIF-1α564组(0.741±0.008)、Ad-HIF-1α564/402组(0.764±0.009)及Ad-HIF-1α564/803组(0.804±0.021),各实验组cTnI表达差异有显著性(P<0.05),其中尤以Ad-HIF-1α564/803组cTnI表达水平最高,与前述其它各组相比差异有显著性(P值均<0.05)。Western blot结果显示:与Ad-LacZ组相比,Ad-HIF-1αnative组、Ad-HIF-1α564组、Ad-HIF-1α564/402组、Ad-HIF-1α564/803组HIF-1α、VEGF及Nkx2.5蛋白均明显升高(P<0.05),其中以Ad-HIF-1α564/402组HIF-1α蛋白表达水平最高(P值均<0.05),以Ad-HIF-1α564/803组VEGF及Nkx2.5蛋白表达水平最高(P<0.05)。结论野生型及突变型HIF-1α基因均可促进hMSCs向心肌细胞分化,且HIF-1α564/803突变型作用最强。Objective To investigate the effects of the native and mutants of hypoxia-inducible factor lot on human mesenchymal stem cells differentiation into cardiomyocyte induced by 5- azacytidine in vitro.Methods To transfect 5-azacytidine induced hMSCs with Ad-LacZ,Ad-HIF- 1αnative ,Ad-HIF-1α564,Ad-HIF-1α564/402 and Ad-HIF-1α564/803 at the optimal multiplicity of infection （MOI） ,cells were harvested at 6 weeks after the transfection,and then the expression of cTnl was detected by ELISA,as well as HIF-1α,VEGF and Nkx2.5 by western blot analysis.Results In contrast, there were more significant differences in expression of cTnl among the control group（ 0.700±0.013 ）, Ad-H IF- 1αnative（ 0.729±0.004）, Ad-HI F-1α564 （ 0.741 ±0.008）, Ad-HIF-1α564/402（0.764±0.009） and Ad-HIF-1α564/803（0.804±0.021 ） （P〈0.05）, and the cTnl expression level transfected by Ad-HIF-1α564/803 was significantly higher as compared with all other experimental groups（all P〈0.05）. Meanwhile,there were also significant differences in expression of HIF-1α,VEGF and Nkx2.5 between Ad-HIF- 1αnative, Ad-HIF- 1α564, Ad- HIF- 1α564/402 and Ad- HIF-1α564/803（P〈0.05） ,and the expression level of HIF-1α was the highest in Ad-HIF-l±564/402 （all P〈0.05）, but the significant higher expression of VEGF and Nkx2.5 were in Ad-HIF-1α564/803 （all P〈0.05,respectively）.Conelusion Both the native and mutants of hypoxia-inducible factor 1α can stably express and promote the effect of hMSC differentiation into cardiomyocytes, specifically enhanced by Ad-HIF-1α564/803.

关 键 词：低氧诱导因子1Α 骨髓间充质干细胞 心肌细胞 5-氮胞苷 分化 

分 类 号：R543[医药卫生—心血管疾病]