黄芪注射液对内皮细胞缺氧复氧损伤的保护作用

Protective Effects of Astragalus Membranaceus Injection on the Endothelial Cells Injured by Hypoxia-Reoxygenation

目的探讨黄芪注射液对内皮细胞缺氧复氧损伤的保护作用及其机制。方法人脐静脉内皮细胞(HUVECs)原代培养 ,传至3代后 ,分为三组 :对照组常规培养 ;缺氧复氧组先缺氧处理1h ,再复氧1h。药物干预组在培养液中加入不同浓度的黄芪注射液 ,12h后进行缺氧复氧处理。各组均测定细胞上清液超氧化物歧化酶 (SOD)活性、丙二醛 (MDA)、一氧化氮 (NO)和内皮素 (ET)含量。结果①缺氧复氧组与对照组比较 ,细胞上清液中SOD活性降低 (P<0.01)、MDA含量升高 (P<0.01) ,NO含量降低 (P<0.01)、ET含量升高 (P<0.01)。②黄芪注射液组与缺氧复氧组比较 ,细胞上清液中SOD活性升高 (P<0.05或P<0.01)、MDA含量降低 (P<0.05或P<0.01) ,在浓度5～40μg/ml范围内该效应与黄芪注射液呈剂量依赖效应 ,但当浓度大于40μg/ml时 ,该效应无明显改变 (P>0.05) ;NO含量升高 (P<0.01)、ET含量降低 (P<0.05或P<0.01) ,直线相关分析显示 :随着黄芪注射液预处理浓度增加 ,培养液中NO的生成量升高 (r1=0.93,P<0.05) ,ET含量降低 (r2= -0.97 ,P<0.05)。结论缺氧复氧过程造成内皮细胞脂质过氧化损伤 ,引起内皮功能紊乱。黄芪注射液可能通过抗脂质过氧化 ,加强对氧自由基的清除 ,对缺氧复氧内皮细胞产生保护作用。

Objective To investigate the protective effect of Astragalus membranaceus injection on the huˉman umbicial vein endothelial cells(hUVECs)injured by hypoxia-reoxygenation and its possible mechanism.Methods hUVECs were cultured and went down to the future generation.After the third generation,experiments were divided into three groups:control group cells were cultured under normal conditions.Cells of hypoxia/reoxyˉgenation group were cultured in closed container with oxygen concentrations below1%for1hour,and then in CO 2 foster box under normal conditions for1hour.hUVECs in astragalus groups were given different dose of Astragalus membranaceus injection at the time of pre-hypoxia,after12hours,they suffered from Hypoxia-Reoxygenation.The activity of superoxide dismutase(SOD),content of malondialdehyde(MDA),endothelin(ET)and nitric oxide(NO)production were determined in cell medium.Results①After treatment with hypoxia/reoxygenation,SOD activity was lower in cell medium(P<0.01),meanwhile the content of MDA became higher(P<0.01).The conˉtent of ETin cell medium became higher(P<0.01),meanwhile NO production was lower in cell medium(32.26±3.88vs87.41±7.63,P<0.01).②After treatment with Astragalus membranaceus injection,SOD activity became higher in cell medium(P<0.05or P<0.01),meanwhile the content of MDA in cell medium was lower(P<0.05or P<0.01),these changes were dose-dependent on the Astragalus membranaceus injection in the range of5～40μg/ml,but they were not significant after dose of astragalus going beyond40μg/ml.The content of ETin cell medium became lower(P<0.05or P<0.01),meanwhile NO production was higher in cell medium(P<0.01),NO production was positively correlated with dose of astragalus(r 1 =0.93,P<0.05),the content of ETwas negˉatively correlated with dose of astragalus(r 2 =-0.97,P<0.05).Conclusions Hypoxia/Reoxygenation could inˉ jure hUVECs correlated with lipid peroxides and lead to endothelial dysfunction.Astragalus membranaceus injection can protect endothelial cells from the injury caused by hypoxia_reoxygenation and its mechanism of action may be related to antioxidant effect of lipids.