原花青素B2保护血管内皮细胞延缓凋亡及机制研究

Study on effect and mechanism of GSPB2 for protecting vascular endothelial cell and delaying apoptosis

目的以H_2O_2诱导人内皮细胞凋亡为模型,观察原花青素B2(GSPB2)延缓人内皮细胞凋亡的保护作用及其机制。方法用0.5mmol H_2O_2预处理细胞,分别加入不同浓度的GSPB2(5.0、10.0、20.0μmol/L)保护细胞,TUNEL法检测细胞凋亡;Western blotting检测凋亡相关分子Caspase-3、Bax、Bcl-2以及影响凋亡的相关分子PI3K、p-Akt、Akt的蛋白表达;Transwell小室检测各组细胞迁移情况。结果 0.5mmol H_2O_2致人内皮细胞凋亡明显增加(P<0.01),GSPB2能明显减轻H_2O_2所致的人内皮细胞凋亡(P<0.05),10μmol/mL的GSPB2就能接近恢复到H_2O_2未处理的对照组水平。进一步研究发现:Caspase-3、bax蛋白表达上调,Bcl-2表达下调;PI3K、p-Akt、Akt表达上调。结论不同浓度的GSPB2对H_2O_2诱导的血管内皮细胞具有保护作用,而且表现为浓度依赖性和时间依赖性,其机制与相关分子PI3K、p-Akt、Akt有关。

Objective To observe the protective effect of GSPB2 for delaying the human endothelial cells apoptosis with H2O2 induced human endothelial cells apoptosis as the model and to investigate its mechanism.Methods The human endothelial cells were pre-treated by 0.5mmol H2O2,then different concentrations of GSPB2（5.0,10.0,20.0μmol/L）was added for protecting cells.The cellular apoptosis was detected by TUNEL method;the apoptosis related molecules,such as Caspase-3,Bax,Bcl-2,and influencing apoptosis related molecules such as PI3 K,p-Akt and Akt protein expression were detected by Western blot;Transwell chamber was used to detect cell migration situation.Results 0.5mmol H2O2 induced obvious increase of human endothelial cells（P〈0.01）.GSPB2 could significantly alleviate the H2O2 induced apoptosis of human endothelial cells（P〈0.05）.10μmol/mL GSPB2 could be close to return to H2O2 untreated control group level.Further found that Caspase-3,Bax protein expression were up-regulaed,Bcl-2expression was down-regulated and the expression of PI3 K,p-Akt and Akt wa sup-regulated.Conclusion The different concentrations of GSPB2 has a protective effect on vascular endothelial cells induced by H2O2,moreover which manifested by the concentration and time dependence.Its mechanism is related with the related molecules PI3 K,p-Akt and Akt