梓葛冻干粉对人脐静脉内皮细胞缺氧/复氧损伤的保护作用  

Protective effect of Zige lyophilized powder on hypoxia/ reoxygenation injury of human umbilical vein endothelial cells

作　　者：张聪聪[1] 马强[1] 薛强[1] 陈刚[2] 徐晓玉[1] 

机构地区：[1]西南大学药学院重庆市药效评价工程技术研究中心,重庆400716 [2]重庆工商大学药物研究中心,重庆400067

出　　处：《中国中药杂志》2012年第7期997-1001,共5页China Journal of Chinese Materia Medica

基　　金：重庆市科技攻关项目(CSTC;2010AC5011);教育部教师科研基金项目(XDJK2009C084;XDJK2010C059);重庆市卫生局中医药科研项目(2010-2-148)

摘　　要：目的:研究中药单体组方梓葛冻干粉对人脐静脉内皮细胞(HUVECs)缺氧/复氧损伤的保护作用。方法:体外培养HUVECs,采用Krebs液建立缺氧/复氧损伤模型。用梓葛冻干粉干预后,采用MTT比色法测定细胞活力、黄嘌呤氧化酶法测定细胞内超氧化物歧化酶(SOD)活性、硫代巴比妥酸显色法测定丙二醛(MDA)含量、硝酸还原酶法测定一氧化氮(NO)含量、2,4-二硝基苯肼显色法测定细胞外乳酸脱氢酶(LDH)释放量;Western blot法分析细胞凋亡相关蛋白Bcl-2,Bax,Caspase-3的表达。结果:与模型组相比,梓葛冻干粉12.5 mg.L-1显著提高细胞活力(P<0.05);梓葛冻干粉49.0,24.5,12.5 mg.L-1均显著提高缺氧/复氧后细胞SOD活性(P<0.05);梓葛冻干粉24.5,12.5 mg.L-1能显著减少LDH的释放量及降低MDA和NO的含量(P<0.05),并上调细胞Bcl-2蛋白表达(P<0.05,P<0.01);梓葛冻干粉49.0,24.5,12.5 mg.L-1显著降低Bax的表达并上调Bcl-2/Bax(P<0.05,P<0.01),且梓葛冻干粉49.0,24.5 mg.L-1显著降低Caspase-3的表达(P<0.01)。结论:梓葛冻干粉可显著拮抗缺氧/复氧对HUVECs的损伤,可能与其抑制细胞过氧化损伤,增强细胞抗氧化和抗凋亡能力有关。Objective： To study the protective effect of single prescription of traditional Chinese medicine Zige lyophilized powder on hypoxia/reoxygenation injury of human umbilical vein endothelial cells（HUVECs）.Method： HUVECs were cultured in vitro and Krebs liquid was adopted to establish the hypoxia/reoxygenation injury model.After intervention of Zige lyophilized powder,MTT colorimetric method was used to determine cell activity,the xanthine oxidase method is adopted to detect the activity of superoxide dismutases（SOD） in cells,the thibabituric acid developing method was adopted to determine the content of malondialdehyde（MDA）,the nitrate reductase method was used to detect the content of nitric oxide（NO）,the 2,4-dinitrophenylhydrazine developing method was adopted to the content of extracellular lactate dehydrogenase（LDH）,and the Western blot method was used to analyze the expression of cell apoptosis-related proteins Bcl-2,Bax and Caspase-3.Result： Compared with the model group,12.5 mg·L-1 of Zige lyophilized powder can enhanced cell viability（P0.05）.Besides,49.0,24.5,12.5 mg·L-1 of Zige lyophilized powder prevents the decline of SOD activity during H/R（P0.05）.Furthermore,24.5,12.5 mg·L-1 Zige lyophilized powder significantly inhibited the increase of MDA and NO content and improved the expression of Bcl-2 protein（P0.05,P0.01）.And 49.0,24.5,12.5 mg·L-1 of Zige lyophilized powder significantly decreased Bax level and up-regulated Bcl-2/Bax ratio.Caspase activation and apoptosis were monitored in the reoxygenated cells.49.0,24.5 mg·L-1 Zige lyophilized powder can decreased Caspase-3 expression（P0.01）.Conclusion： Zige lyophilized powder can prevent H/R induced injury to HUVECs,which may be related to its effect in inhibiting cell peroxidation injury and enhancing cell antioxidant and antiapoptotic capacities

关 键 词：梓葛冻干粉 缺氧/复氧损伤 人脐静脉内皮细胞 

分 类 号：R285.5[医药卫生—中药学]