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中国应用生理学杂志 ›› 2018, Vol. 34 ›› Issue (2): 164-168.doi: 10.12047/j.cjap.5659.2018.040

• 研究论文 • 上一篇    下一篇

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

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   

  1. 天津医科大学基础医学院药理学系, 天津 300070
  • 收稿日期:2017-12-11 修回日期:2018-01-17 出版日期:2018-03-28 发布日期:2018-05-22
  • 通讯作者: 宋君秋, 刘艳霞 E-mail:liuyanxia126@126.com;songjunqiu@126.com
  • 基金资助:
    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)

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

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   

  1. Department of Pharmacology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China
  • Received:2017-12-11 Revised:2018-01-17 Online:2018-03-28 Published:2018-05-22
  • Supported by:
    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12 h/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,AST 20、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/REMVs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p-ERK1/2蛋白质水平有关。

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

Abstract: 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 10μg/ml; different doses of AST groups:thoracic aortic rings of rats were treated with 10, 20, 40, 60 mg/L AST co-incubated with 10μg/ml H/R-EMVs respectively; control group were treated with the same volume of D-Hank'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:Tenμg/ml H/R-EMVs could impaire the relaxation of rat thoracic aortic rings significantly (P<0.01). Compared with H/R-EMVs 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 with AST (P<0.01). Conclusion:AST could effectively ameliorate endotheliumdependent relaxation of rat thoracic aortic rings impaired by H/R-EMVs in a concentration-dependent manner, the mechanism might involve the increase in production of NO, and the protein level of p-eNOS, p-Akt and p-ERK1/2.

Key words: Astragaloside IV, microvesicles, hypoxia/reoxygenation, HUVECs, aortic rings, rats

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