黄芪苷Ⅳ对微囊泡损伤的大鼠胸主动脉环舒张功能的保护作用  

The protective effects of Astragaloside Ⅳ on diastolic function of rat thoracic aortic rings impaired by microvesicles

作　　者：李烨仪 尚曼 张琨玮 韦苏 刘超 祝倩 赵俊玉 吴艳娜 宋君秋 刘艳霞 LI Ye-yi;SHANG Man;ZHANG Kun-wei;WEI Su;LIU Chao;ZHU Qian;ZHAO Jun-yu;WU Yan-na;SONG Jun-qiu;LIU Yan-xia(Department of Pharmacology,School of Basic Medical Sciences,Tianjin Medical University,Tianjin 300070,Chin)

机构地区：[1]天津医科大学基础医学院药理学系,天津300070

出　　处：《中国应用生理学杂志》2018年第2期164-168,共5页Chinese Journal of Applied Physiology

基　　金：天津市自然科学基金项目(11JCZDJC18300);天津市高等学校科技发展基金计划项目(20110106)

摘　　要：目的:以缺氧/复氧诱导人脐静脉内皮细胞(HUVECs)释放的微囊泡(H/R-EMVs)处理大鼠胸主动脉环,造成其舒张功能损伤,探究黄芪苷Ⅳ(AST)对大鼠胸主动脉环舒张功能的影响及相关机制。方法:采用缺氧12h/复氧4 h的方法诱导体外培养的HUVECs产生MVs,H/R-EMVs保存于D-Hank’s液中备用。雄性Wistar大鼠开胸取出胸主动脉,制备3~4 mm宽、内皮完整的胸主动脉环。实验分为6组:H/R-EMVs组,在孵育胸主动脉环的培养基中加入H/R-EMVs,使其终浓度为10μg/ml;不同剂量AST组分别采用10、20、40、60 mg/L AST与10μg/ml H/R-EMVs共同孵育胸主动脉环;对照组给予等体积的D-Hank’s溶液。孵育时间为4 h,每组各测定5个血管环。观察AST对舒张功能的影响,检测一氧化氮(NO)含量及t-eNOS、p-eNOS、t-Akt、p-Akt、ERK1/2和p-ERK1/2蛋白质水平。结果:H/R-EMVs对大鼠胸主动脉环舒张功能有明显的抑制作用(P<0.01)。与H/R-EMVs组相比,AST20、40和60 mg/L组剂量依赖性地提高大鼠胸主动脉环的舒张率(P<0.01),使NO含量增加(P<0.05,P<0.01);t-eNOS、t-Akt和ERK1/2蛋白质水平不变,p-eNOS、p-Akt和p-ERK1/2蛋白质水平增高(P<0.01)。结论:AST可显著改善H/R-EMVs损伤的大鼠胸主动脉环的舒张功能,其机制与提高NO含量及增加p-eNOS、p-Akt和pERK1/2蛋白质水平有关。Objective： To investigate the effects of Astragaloside IV（ AST） on diastolic function of rat thoracic aorta rings which was injured by microvesicles derived from hypoxia/reoxygenation（ H/R）-treated human umbilical vein endothelial cells（ HUVECs）,and the mechanism of AST. Methods： H/R-induced endothelial microvesicles（ H/R-EMVs） were generated from cultured HUVECs in vitro under the condition of hypoxia for 12 hour/Reoxygenation for 4 hour,H/R-EMVs were stored in D-Hank＇s solution. Male Wistar rats were underwent thoracotomy,the thoracic aorta with intact endothelium were carefully removed and cut into 3 ～ 4 mm rings. The experiment was divided into six groups. H/R-EMVs group： thoracic aortic rings of rats were incubated in culture medium and treated with H/R-EMVs in a final concentration of 10 μg/ml; different doses of AST groups： thoracic aortic rings of rats were treated with 10,20,40,60 mg/L AST co-incubated with 10 μg/ml H/R-EMVs respectively; control group were treated with the same volume of DHank＇s solution. Duration of incubation was 4 h,each group was tested in five replicate aortic rings. Effects of AST on endothelium-dependent relaxation were detected. The production of nitric oxide（ NO） and the level of endothelial NO synthase（ eNOS）,phosphorylated eNOS（ p-eNOS,Ser-1177）,serine/threonine kinase（ Akt）,phosphorylated Akt（ p-Akt,Ser-473）,extracellular regulated protein kinases（ ERK1/2） and phosphorylated ERK1/2（ p-ERK1/2,Thr202/Tyr204） of rat thoracic aortic rings were detected. Results： Ten μg/ml H/R-EMVs could impaire the relaxation of rat thoracic aortic rings significantly（ P〈0. 01）. Compared with H/REMVs group,relaxation of rat thoracic aortic rings was increased by 20,40 and 60 mg/L AST in a concentration-dependent manner（ P〈0. 01）,the level of NO production was also enhanced（ P〈0. 05,P〈0. 01）. The level of t-eNOS,t-Akt and ERK1/2 was not changed,but the level of p-eNOS,p-Akt and p-ERK1/2 increased by the treatment w

关 键 词：黄芪苷Ⅳ 微囊泡 缺氧/复氧 人脐静脉内皮细胞 胸主动脉环 大鼠 

分 类 号：R73-3[医药卫生—肿瘤]