甘肃黄芪黄酮对AngⅡ致内皮细胞凋亡的保护作用

目的探讨甘肃黄芪黄酮化合物芒柄花素与毛蕊异黄酮-7-O-β-D-吡喃葡萄糖对血管紧张素Ⅱ(AngⅡ)诱导人脐静脉内皮细胞(HUVECs)凋亡的影响。方法将不同浓度上述两种黄酮单体(0.01、0.1、1.0、10mg.L-1)分别与AngⅡ(10-6mol.L-1)和HUVECs作用24h后,MTT比色法检测细胞存活率,Hoechst33258荧光染色法观察细胞核形态变化,流式细胞仪及激光共聚焦仪检测细胞凋亡,并观察凋亡基因Fas表达的变化。结果甘肃黄芪黄酮化合物芒柄花素与毛蕊异黄酮-7-O-β-D-吡喃葡萄糖呈剂量依赖性地抑制AngⅡ引起的细胞存活率下降(P<0.05),AngⅡ(10-6mol.L-1)孵育24h后,内皮细胞凋亡率明显多于对照组(P<0.01);芒柄花素(1.0mg.L-1)与毛蕊异黄酮-7-O-β-D-吡喃葡萄糖(1.0mg.L-1)可抑制AngⅡ(10-6mol.L-1)诱导的内皮细胞凋亡(P<0.01),并减少Fas的表达(P<0.05);以上实验芒柄花素与毛蕊异黄酮-7-O-β-D-吡喃葡萄糖两化合物间比较无差异(P>0.05)。结论甘肃黄芪黄酮化合物可减轻AngⅡ诱导的HUVECs凋亡,其机制之一可能与其降低Fas的表达有关。

Protective effect of astragalus flavone of Gansu on endothelial cells apoptosis induced by angiotensin Ⅱ in culture

Aim To study protective effects by flavones(formononetin and calycosin-7-O-β-D-glucopyranoside)of Radix astragali from Gansu province on angiotensin II(AngII)-induced the injury of human umbilical vein endothelial cells(HUVECs)in vitro.Methods Eight groups of cultured HUVECs were incubated with AngII(10-6 mol·L-1)and Astragalus flavone for 24 h,which were formononetin(0.01、0.1、1.0、10 mg·L-1)and calycosin-7-O-β-D-glucopyranoside(0.01、0.1、1.0、10 mg·L-1).The viability of endothelial cells was monitored by MTT colorimetric assay.Hoechst 33258 dyeing was used to observe AngII-induced nuclear morphological changes.The apoptosis of HUVECs was detected by flow cytometry and Laser-Scanning Confocal Microscope,also the change of Fas expression was observed.Results The formononetin and calycosin-7-O-β-D-glucopyranoside could inhibit the cells viability induced by AngII in a concentration-dependent manner(P<0.05).It was found that incubation with AngII increased the HUVECs' apoptosis(P<0.01).The formononetin(1.0 mg·L-1)and calycosin-7-O-β-D-glucopyranoside(1.0 mg·L-1)significantly decreased Ang Ⅱ-induced HUVECs' apoptosis(P<0.01)and the expression of Fas(P<0.05).There were no significant differences between for-mononetin and calycosin-7-O-β-D-glucopyranoside groups in above experiments(P>0.05).Conclusion Two flavones separated from Astragalus in Gansu province have precisely protective effects on HUVECs via inhibition of AngⅡ induced apoptosis and one of mechanisms of their action may be related to alleviate the expression of Fas.