突变型低氧诱导因子1α腺病毒载体对骨髓间充质干细胞增殖的影响

发布时间：2018-05-26 01:20

本文选题：干细胞 + 间质干细胞　；参考：《中国组织工程研究》2017年25期

【摘要】：背景:目前发现低氧诱导因子1α(hypoxia inducible factor 1,HIF1α)可提高组织和细胞在缺血环境下继续生存的能力,在促进血管再生方面有重要意义。目的:构建三点突变型HIF1α联合示踪因子hrGFP双基因重组腺病毒载体Ad-CMV-HIF1α~(mu)-IRES-hrGFP-1,研究其对骨髓间充质干细胞增殖的影响。方法:将第3代骨髓间充质干细胞分组培养,实验组以突变基因Ad-CMV-HIF1α~(mu)-IRES-hrGFP-1病毒液转染细胞,阳性对照组以未突变基因Ad-CMV-HIF1α-IRES-hrGFP-1病毒液转染细胞,阴性对照组以Ad-CMV-IRES-hrGFP-1空病毒液转染细胞,空白对照组不转染任何病毒液。转染72 h后,进行HIF1α基因mRNA及蛋白检测;转染24,48,72,96,120 h,MTT法检测细胞增殖。结果与结论:(1)HIF1α基因mRNA检测:实验组、阳性对照组HIF1α基因mRNA表达高于阴性对照组、空白对照组(P0.05),其余组间两两比较差异无显著性意义;(2)HIF1α蛋白检测:实验组细胞中HIF1α蛋白表达高于阳性对照组、阴性对照组、空白对照组(P0.05),后3组间比较差异无显著性意义;(3)细胞增殖:阳性对照组与空白对照组细胞生长速度均较慢,两组间相比较无显著性意义;实验组细胞增殖程度高于阳性对照组、空白对照组(P0.05);(4)结果表明,重组腺病毒介导的三点突变型HIF1α联合hrGFP基因转染骨髓间充质干细胞后,不仅可在常氧条件下持续表达目的蛋白HIF1α,同时也可促进骨髓间充质干细胞的增殖。
[Abstract]:Background: at present, it is found that hypoxia inducible factor 1 alpha (hypoxia inducible factor 1, HIF1 a) can improve the ability of tissue and cells to continue to survive in the ischemic environment, and it is of great significance in promoting vascular regeneration. Objective: to construct the three point mutant HIF1 alpha combined with hrGFP double gene recombinant adenovirus vector Ad-CMV-HIF1 a ~ (MU) -IRES-hrGFP-. 1, study its effect on the proliferation of bone marrow mesenchymal stem cells. Methods: the third generation bone marrow mesenchymal stem cells were grouped in groups. The experimental group transfected cells with the mutant gene Ad-CMV-HIF1 alpha ~ (MU) -IRES-hrGFP-1 virus solution. The positive control group transfected the cells with the Ad-CMV-HIF1 alpha -IRES-hrGFP-1 virus of the non mutant gene, and the negative control group was Ad-CMV-IRES-hrG FP-1 empty virus fluid transfected cells, blank control group did not transfect any virus fluid. After transfection of 72 h, HIF1 alpha gene mRNA and protein detection, 24,48,72,96120 h, MTT method to detect cell proliferation. Results and conclusion: (1) HIF1 a gene mRNA detection: experimental group, mRNA control group HIF1 alpha gene mRNA higher than the negative control group, the blank control group (P0) .05), there was no significant difference between the 22 other groups. (2) HIF1 alpha protein detection: the expression of HIF1 alpha protein in the experimental group was higher than that in the positive control group, the negative control group and the blank control group (P0.05), and there was no significant difference between the 3 groups. (3) the growth rate of the cells in the positive control group and the blank control group were both slower and two groups. The proliferation of cells in the experimental group was higher than that in the positive control group and the blank control group (P0.05). (4) the results showed that the recombinant adenovirus mediated three point mutant HIF1 alpha combined with hrGFP gene transfected to bone marrow mesenchymal stem cells could not only express the target protein HIF1 alpha continuously under the normal oxygen strip, but also promote the bone marrow. Proliferation of mesenchymal stem cells.
【作者单位】：锦州医科大学附属第一医院整形外科;锦州医科大学附属第一医院临床生物样本中心;
【基金】：辽宁医学院校长基金——临床医学建设专项基金项目资助(XZJJ20140215)~~
【分类号】：R329.2

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