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目的探讨补阳还五汤抑制Hcy诱导的内皮细胞凋亡与MAPK/ERK途径的关系。方法内皮细胞用Hcy(2mmol/L)作用时间为24h诱导损伤,治疗组给予补阳还五汤含药血清(5%、10%,20%),SB203580,PD98059,24 h后,采用Annexin V-FITC和PI双染法,用流式细胞术检测内皮细胞凋亡率,蛋白质印迹技术检测Caspase-3、Caspase-9,MAPK、ERK的蛋白质水平。结果 Hcy组HUVEC凋亡明显(P<0.01),与空白对照组相比Caspase-3,Caspase-9,MAPK,ERK蛋白含量明显升高,细胞凋亡率检测3个剂量的BYHW显著抑制Hcy诱导的内皮细胞凋亡,不同剂量的补阳还五汤显著降低内皮细胞中Caspase-3、Caspase-9,MAPK,ERK蛋白表达。结论补阳还五汤能显著抑制Hcy诱导的内皮细胞的凋亡,其可能的机制是调控内皮细胞中的MAPK/ERK信号通路的表达有关。
Objective To investigate the relationship between Buyang Huanwu decoction and Hcy-induced endothelial cell apoptosis and MAPK / ERK pathway. Methods Endothelial cells were induced by Hcy (2mmol / L) for 24 hours. The treatment group was given Fuyang Huanwu Decoction (5%, 10%, 20%), SB203580 and PD98059 for 24 h, The expression of Caspase-3, Caspase-9, MAPK and ERK were detected by Western blotting. The apoptosis rate of endothelial cells was detected by FITC and PI double staining. Results The apoptosis of HUVEC in Hcy group was significantly higher than that in control group (P <0.01). Compared with the blank control group, the protein levels of Caspase-3, Caspase-9, MAPK and ERK were significantly increased. The endothelial cells were treated with Buyang Huanwu Decoction at different doses for a dose-dependent manner. The expressions of Caspase-3, Caspase-9, MAPK and ERK in endothelial cells were significantly decreased. Conclusion Buyang Huanwu decoction can significantly inhibit Hcy-induced endothelial cell apoptosis, and its possible mechanism is to regulate the expression of MAPK / ERK signaling pathway in endothelial cells.