摘要
[目的]探讨缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)对间充质干细胞基质细胞衍生因子-1(stromal cell-derived factor-1,SDF-1)/CXC趋化因子受体4(chemokine receptor 4,CXCR4)的调控作用。[方法]贴壁法培养ICR小鼠骨髓间充质干细胞(mesenchymal stem cells,MSCs);采用RNA干扰技术用SuperFectinTMⅡ转染HIF-1αsiRNA于MSCs;实验分为五组:常氧组、缺氧组、转染Control siRNA组、转染HIF-1αsiRNA常氧培养组、转染HIF-1αsiRNA缺氧培养组;RT-PCR检测HIF-1α、SDF-1α、CXCR4 mRNA表达水平;Western blotting检测HIF-1α、SDF-1α、CXCR4蛋白表达水平。[结果]同常氧组比较,缺氧组HIF-1α、SDF-1α、CXCR4 mRNA及蛋白表达水平提高(P<0.05);同转染Control siRNA组比较,转染HIF-1αsiRNA常氧培养组HIF-1α、SDF-1α、CXCR4 mRNA及蛋白表达水平降低(P<0.05);同缺氧组比较,转染HIF-1αsiRNA缺氧培养组HIF-1α、SDF-1α、CXCR4 mRNA及蛋白表达水平降低(P<0.05)。[结论]缺氧可使MSCs的HIF-1α、SDF-1α、CXCR4的表达提高,常氧与缺氧条件下抑制HIF-1α的表达可使SDF-1α、CXCR4的表达降低,HIF-1α在间充质干细胞中对SDF-1/CXCR4有调控作用。
[Aim] To study the regulation of HIF-1α for SDF-1/CXCR4 in MSCs. [Methods] MSCs were cultured by adherence method. HIF-1αsiRNA was transfected to MSCs by SuperFectinTM Ⅱ. The experiment was divided into five groups, normoxia group,hypoxia group,control siRNA transfected group, HIF-1α siRNA transfected normoxic group, HIF-1α siRNA transfected hypoxia group. Reverse transcdption-polymerase chain reaction was utilized to determine HIF-1α, SDF-1α, CXCR4 mRNA expression in MSCs.Western blotting was employed to detect HIF-1α, SDF-1α, CXCR4 protein expression in MSCs. [Result] Compared with the normoxia group, HIF-1α, SDF-1α and CXCR4 mRNA and protein expression increased in the hypoxia group（P〈0.05）. Compared with the control siRNA transfected group, HIF-1α, SDF-1α and CXCR4 mRNA and protein expression decreased in the HIF-1α siRNA transfected normoxic group（P〈0.05）; Compared with the hypoxia group, HIF-1α, SDF-1α and CXCR4 mRNA and protein expression decreased in the HIF-1α siRNA transfected hypoxia group（P〈0.05）. [Conclusion] These results verified that the expressions of HIF-1α, SDF-1αand CXCR4 were increased in hypoxic environment. Inhibition of HIF-1αexpression decreased SDF-1αand CXCR4 expression both in normoxia and hypoxia. HIF-1αplays an important role in regulating SDF-1/CXCR4 in mesenchymal stem cells.
出处
《浙江中医药大学学报》
CAS
2014年第3期237-242,254,共7页
Journal of Zhejiang Chinese Medical University
基金
浙江省自然科学基金(LY12C10001)
国家自然科学基金(81270566)~~
