丁苯酞对缺氧缺糖条件下血管内皮细胞VEGF和HIF-1α表达的影响

摘要

AIM : To investigate the mechanism that dl - 3 - n - butylphthalide ( NBP) protects cells from the induced by oxygen - glucose deprivation ( OGD) .METHODS : Human umbilical vein endothelial cells ( HUVECs) were exposed to OGD to induce endothelial damage.Endothelial injury was assessed by measuring the changes of chromatin morphology and MTT method.The protein levels of vascular endothelial growth factor ( VECF) and hypoxia inducible factor I alpha ( HIF - 1α) were determined by immunofluorescence and quantitatively analyzed with the software IPP.Western blotting was applied to verify the results.RESULTS : NBP at the concentrations of 0.01 to 100 μmol/L dose - depenclently protected against OGD - induced cell damage.Compared with OGD group, NBP enhanced OGD - induced the expression of VEGF and HIF - 1α , and the difference was statistically significant.The expression of VEGF and HIF - 1α reached to the peak at the time points of 6 h and 8 h after OGD, respectively.CONCLUSION: Under the condition of OGD, NBP enhances the expression of HIF - 1α in HUVECs, subsequently promotes the expression of downstream VEGF, and eventually elevates the survival of the cells.%目的:探讨丁苯酞(NBP)保护缺氧缺糖(OGD)细胞损伤的机制.方法:对人脐静脉内皮细胞(HUVECs)予以OGD损伤,MTT法检测细胞活性;Hoechst 33342染色检测细胞核形态;免疫荧光法观察血管内皮生长因子(VEGF)和缺氧诱导因子-1α(HIF-1α)表达,IPP软件分析荧光强度,进行定量分析;Western blotting检测加以验证.结果:NBP在0.01- 10 μmol/L浓度之间剂量依赖性减少了OGD导致的细胞活性下降和细胞核形态改变.NBP促进了OGD后HUVECs内VEGF和HIF-1α的表达,均高于OGD组,荧光定量分析差异显著.两者表达的高峰分别为OGD后6 h和8 h.结论:NBP可以促进内皮细胞在缺氧缺糖条件下HIF-1α的表达,从而引起下游VEGF的表达增多,最终保护内皮细胞免受缺氧缺糖损伤.