【摘要】 目的:探讨麦冬皂苷D对氧糖剥夺/复氧(OGD/R)后星形胶质细胞HIF-1α-VEGF通路的作用。方法:分离培养和鉴定新生大鼠星形胶质细胞,建立星形胶质细胞OGD/R模型;设正常组、模型组、麦冬皂苷D给药组;在OGD4h/R2h、4h、6h后用ELISA检测星形胶质细胞细胞培养液中VEGF蛋白表达量,Western Blot检测星形胶质细胞胞核内HIF-1α蛋白的含量,免疫荧光检测星形胶质细胞及其在OGD/R后细胞的光密度、面密度和阳性细胞数量。结果:模型组VEGF、HIF-1α蛋白含量高于正常组(P<0.05,P<0.01),麦冬皂苷D给药组高于模型组(P<0.05,P<0.01),且随再灌注时间延长,VEGF、HIF-1α蛋白含量呈增加趋势。模型组星形胶质细胞的面密度、光密度、阳性细胞个数较正常组均明显降低(P<0.01),麦冬皂苷D给药组星形胶质细胞的面密度、光密度、阳性细胞个数较模型组均显著升高(P<0.05,P<0.01)。结论:麦冬皂苷D能够促进OGD/R后新生鼠离体星形胶质细胞的增殖;麦冬皂苷D能够明显上调OGD/R后新生鼠离体星形胶质细胞VEGF、HIF-1α的蛋白含量;麦冬皂苷对OGD/R后神经损伤的保护及修复作用可能是通过星形胶质细胞HIF-1α-VEGF通路实现的。更多还原

【Abstract】 Objective: To discuss the effects of ophiopogonin D on the HIF-1α-VEGF path of neonatal rat cerebral cortex astrocytes in vitro after oxygen glucose deprivation and reperfusion. Methods: Isolated,cultured and identified neonatal rat astrocytes, established the OGD/R model of neonatal rat astrocytes in vitro; set up normal group, model group, and ophiopogonin D treatment group, after oxygen glucose deprivation 4 h and reperfusion 2 h, 4 h and 6 h, analyzed separately the expression levels of VEGF protein in the culture supernatant of astrocytes by ELISA, HIF-1α protein in the nucleus of astrocytes by Western Blot, and the optical density, areal density and the number of positive cells of OGD/R astrocytes by double-labelling immunofluorescence. Results: The content of VEGF and HIF-1α in model group was higher than the normal(P<0.05, P<0.01), while ophiopogonin D treatment group was higher than the model(P<0.05, P<0.01), following reperfusion time lasted, both the expression levels of VEGF and HIF-1α appeared upward trend. Corresponding to this, all of the optical density, areal density and the number of positive cells of astrocytes in model group obviously decreased compared with the normal(P<0.01), and ophiopogonin D treatment group remarkably increased compared with the model, differences of which had statistical significance. Conclusion: ophiopogonin D could promote the proliferation of neonatal rat cerebral cortex astrocytes in vitro after OGD/R. Ophiopogonin D could raise up the expression levels of VEGF and HIF-1α of neonatal rat cerebral cortex astrocytes in vitro after OGD/R; the protective and adjective effects of ophiopogonin D on nerve injury after oxygen glucose deprivation and reperfusion achieved potentially though HIF-1α-VEGF path of neonatal rat cerebral cortex astrocytes in vitro.更多还原