蛇床子素通过Akt/eNOS降低氧化低密度脂蛋白诱导血管内皮细胞凋亡

Protective effects of osthole on ox-LDL-induced-apoptosis of human umbilical vein endothelial cells via Akt/eNOS

目的:探讨蛇床子素对低密度脂蛋白诱导的人脐静脉血管内皮细胞(HUVEC)凋亡的影响及其机制研究。方法:采用HUVEC细胞,设置正常组、模型组和给药组,细胞培养12 h后,低密度脂蛋白进行细胞损伤,同时给予不同浓度蛇床子素(10、20、40μmol/L)进行干预,应用MTT法检测细胞活力;Annexin V-FITC标记的流式细胞术检测细胞凋亡;检测一氧化氮生成量;免疫印迹法检测p-AKt和p-eN OS蛋白表达水平。结果:与正常组相比,模型组细胞活力和凋亡率有显著差异;与模型组相比,蛇床子素提高细胞活力,抑制细胞细胞凋亡,作用效果呈剂量依赖性,20、40μmol/L蛇床子素均有显著性差异;与模型组相比,20、40μmol/L蛇床子素显著促进细胞一氧化氮合成;蛋白质检测结果显示,与模型组相比,蛇床子素可上调HUVEC细胞中Akt和eN OS蛋白磷酸化水平。结论:蛇床子素能够抑制ox-LDL诱导HUVEC的凋亡而发挥其保护作用,其作用机制可能与其激活Akt/eNOS/NO的信号通路相关。

Objective： Study the protective effect and mechanism of osthole on ox-LDL-induced apoptosis of human umbilical vein endothelial cells（ HUVEC）.Methods： HUVEC cells were cultured and produced the mode of apoptosis.Cell groups： HUVEC cells were divided into control group（ Control）,model group（ ox-LDL）,osthole group（ Osthole ＋ ox-LDL）.MTT assay was used to detect cell viability and determine the optimal effectively protect concentration of osthole; Apoptotic rate was measured by flow cytometry; The secretary content of nitric oxide（ NO） was detected by the commercial reagents; Western Blot assay was performed to detect the expressions of p-Akt and p-e NOS protein.Results： Osthole（ 20,40 μmol/L） treatment dramatically inhibited HUVEC apoptosis compared to model group.NO contents in osthole groups（ 20,40 μmol/L） were also significantly different compared with those in model group.In osthole groups（ 20、40 μmol/L） the expressions of p-Akt and p-e NOS protein were significantly increased.Conclusion： Osthole has inhibition effect on HUVEC apoptosis,the osthole can protect HUVEC damaged by ox-LDL,which plays a role via Ak T/eNOS/NO pathway.