黄芪多糖减轻氯化钴致人肺动脉内皮细胞损伤  

Astragalus polysaccharide alleviates injury of human pulmonary arterial endothelial cells induced by cobalt chloride

作　　者：张甲翠[1] 魏海东[3] 王海燕 应佳丽[2] ZHANG Jiacui;WEI Haidong;WANG Haiyan(Department of Respiratory,People's Hospital of Gansu Province,Lanzhou 730000,CHINA)

机构地区：[1]甘肃省人民医院呼吸科,甘肃兰州730000 [2]甘肃省人民医院中医科,甘肃兰州730000 [3]兰州大学第二附属医院呼吸科 [4]和政县新营卫生院全科

出　　处：《江苏医药》2020年第6期576-580,共5页Jiangsu Medical Journal

基　　金：甘肃省卫生行业科研计划项目(GSWSKY-2015-02);甘肃省人民医院院内科研基金项目(18GSSY4-26)。

摘　　要：目的探讨黄芪多糖(APS)对氯化钴(CoCl2)致人肺动脉内皮细胞(HPAEC)损伤的保护作用及其可能机制。方法使用不同浓度CoCl2 100、150、200、250μmol/L诱导HPAEC低氧,采用MTT法检测细胞活性,Western blot法检测低氧诱导因子1α(HIF-1α)和Bcl-2蛋白表达。随后用不同浓度APS 100、200、400μmol/L干预,采用RT-PCR和Western blot法分别检测核因子E2相关因子2(NRF2)、SOD、谷胱甘肽巯基转移酶(GST)mRNA和蛋白表达。结果 CoCl2处理24、48 h时,HPAEC存活率呈浓度依赖性地降低(P<0.05),而HIF-1α和Bcl-2蛋白表达呈浓度依赖性地增加(P<0.05)。随着APS浓度增加,GST和SOD mRNA和蛋白表达逐渐增加(P<0.05),而NRF2 mRNA和蛋白表达逐渐减少(P<0.05)。此外,使用APS 200μmol/L干预后,HIF-1α和Bcl-2蛋白表达呈时间依赖性地减少(P<0.05)。结论 APS可抑制CoCl2所致HPAEC低氧时NRF2活性以及HIF-1α和Bcl-2蛋白表达,增强SOD和GST多种抗氧化酶的表达,从而减轻HPAEC内氧化应激损伤。Objective To investigate the protective effect and underlying mechanism of astragalus polysaccharide(APS) on the injury of human pulmonary arterial endothelial cells(HPAEC) induced by cobalt chloride(CoCl2).Methods The hypoxia of HPAEC was induced by different concentrations of CoCl2 100,150,200 and 250 μmol/L.The cell viability was measured by MTT assay.The protein expressions of hypoxia-inducible factor-1α(HIF-1α) and Bcl-2 were determined by Western blot.Then after the intervention of different concentrations of APS 100,200 and 400 μmol/L,the mRNA and protein expressions of nuclear factor erythroid-2 related factor 2(NRF2),SOD and glutathione S-transferase(GST) were detected by RT-PCR and Western blot,respectively.Results The survival rate of HPAEC was decreased in a concentration-dependent manner(P<0.05),while the protein expressions of HIF-1α and Bcl-2 were increased in a concentration-dependent manner at 24 and 48 hours after the treatment of CoCl2(P<0.05).With the increase of APS,the mRNA and protein expressions of GST and SOD were gradually increased(P<0.05),while which of NRF2 were gradually decreased(P<0.05).Moreover,APS 200 μmol/L inhibited the protein expressions of HIF-1α and Bcl-2 in a time-dependent manner(P<0.05).Conclusion APS could inhibit the activity of NRF2 and expressions of HIF-1α and Bcl-2 in the hypoxia model of HPAEC induced by CoCl2,enhance the expression of antioxidant enzymes SOD and GST,and alleviate the oxidative stress injury in HPAEC.

关 键 词：黄芪多糖 氯化钴 低氧诱导因子1Α B细胞淋巴瘤-白血病2 人肺动脉内皮细胞 

分 类 号：R563[医药卫生—呼吸系统]