microRNA-132在人脐静脉内皮细胞中调控作用的研究

Effect of regulation in human umbilical vein endothelia cells treated by microRNA-132

摘要:

目的：探讨miR-132在人脐静脉内皮细胞(human umbilical vein endothelia cells，HUVEC)中的调控作用。

方法：体外低氧培养人脐静脉内皮细胞6h后继续常氧培养3、6、12、24h，利用Real-time PCR检测miR-132和PGC-1α mRNA表达变化，Western-blot检测PGC-1α 蛋白表达变化，及观察各组与正常培养的细胞之间的表达差异。通过对人脐静脉内皮细胞转染miR-132模拟物与拮抗剂后，再分别置于常氧和低氧环境中培养，利用Real-time PCR检测不同氧条件下miR-132和PGC-1α mRNA的表达变化，Western-blot检测PGC-1α蛋白表达变化。

结果：miR-132和PGC-1α在细胞低氧培养后继续常氧培养3h时蛋白与mRNA表达量最高，与正常培养的细胞表达差异最为明显(P<0.01)。细胞转染后可观察到miR-132和PGC-1α在低氧组表达量要高于常氧组，而两组中转染miR-132模拟物后的表达量要高于转染拮抗剂组(P<0.01)。

结论：miR-132在低氧时的人脐静脉内皮细胞中表达明显上调，对PGC-1α存在调控作用。

Abstract:

AIM: To evaluate the regulatory effect of microRNA-132(miR-132)in human umbilical vein endothelial cell(HUVEC).

METHODS: In vitro cultured human umbilical vein endothelia cells in hypoxic environment for 6h, then maintained under normal oxygen condition for 3h, 6h, 12h, 24h. miR-132 and peroxisome-proliferator-activated receptor-γ coactivator-1α(PGC-1α)expression was detected by quantitative Real-time polymerase chain reaction and Western blot analysis. Human umbilical vein endothelial cells transfected miR-132 mimic and miR-132 inhibitor(anti-miR-132)were measured by quantitative Real-time polymerase chain reaction and Western blot.

RESULTS: miR-132 and PGC-1α expression was significantly(P<0.01)upregulated in the hypoxic environment of cells at 3h compared with the normal oxygen condition. After cells transfection, the hypoxic environment the miR-132 and PGC-1α expression were markedly increased compared with the normal oxygen condition. The cells transfected miR-132-mimic, the expression of the miR-132 and PGC-1α were higher than that of transfected anti-miR-132 and contrast group(P<0.01).

CONCLUSION: miR-132 level is highly expressed in the HUVEC under hypoxia and may be an effect of regulation for PGC-1α.