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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (5): 390-393.doi: 10.12047/j.cjap.5896.2020.083

• 研究论文 • 上一篇    下一篇

Apelin-13对LPS诱导人脐静脉内皮细胞屏障损伤的干预作用*

林道朋1, 陈莎2, 陆溧玲1, 王玉3, 吕芳芳1, 施林微1, 孔晓霞3, 单小鸥1, 张海邻1△   

  1. 1.温州医科大学附属第二医院育英儿童医院, 浙江 温州 325035;
    2.鄂东医疗集团黄石市中心医院湖北理工学院附属医院病理科, 湖北 黄石 435000;
    3.温州医科大学基础医学院低氧研究所, 浙江 温州 325035
  • 收稿日期:2019-06-19 修回日期:2020-06-24 发布日期:2021-02-25
  • 通讯作者: Tel: 0577-86689746; E-mail: zhlwz97@hotmail.com
  • 基金资助:
    *重大新药创制国家科技重大专项(2020ZX09201002-005);国家自然科学基金资助项目(81973382);浙江省自然科学基金资助项目(LY17H010009,LY17H010005)

Effects of Apelin-13 on barrier injury of human umbilical vein endothelial cells induced by LPS

LIN Dao-peng1, CHEN Sha2, LU Li-ling1, WANG Yu3, LYU Fang-fang1, SHI Lin-wei1, KONG Xiao-xia3, SHAN Xiao-ou1, ZHANG Hai-lin1△   

  1. 1. The Second Affiliated Hospital and Yuying Children's Hospital, Wenzhou Medical University, Wenzhou 325035;
    2. Department of Clinical Laboratory, Huangshi Central Hospital of Erdong Medical Group Affiliated Hospital of Hubei Polytechnic University, Huangshi 435000;
    3. School of Basic Medicine, Institute of Hypoxia Research, Wenzhou Medical University, Wenzhou 325035, China
  • Received:2019-06-19 Revised:2020-06-24 Published:2021-02-25

摘要: 目的: 探讨Apelin-13对LPS诱导人脐静脉内皮细胞(HUVECs)屏障损伤的影响。方法: 将体外培养的HUVECs分为4组:正常对照组、LPS组、Apelin-13+LPS组、Apelin-13组。用5 μg/ml LPS作用细胞24 h, 复制屏障功能受损模型。1 μmol/L Apelin-13提前30 min给予,再给予LPS作用24 h,确定Apelin-13的影响。通过CCK8法检测Apelin-13对细胞活力的影响;Western blot检测血管内皮细胞钙粘蛋白(VE-cadherin)、纤维状肌动蛋白(F-actin)表达变化;免疫荧光检测VE-cadherin、F-actin的表达变化及核转录因子Kappa B(NF-κB p65)入核情况。结果: 与正常对照组相比,单独给予Apelin-13对细胞活力无明显影响。与正常对照组相比,LPS组细胞活力明显下降(P<0.01),VE-cadherin蛋白表达下降(P<0.01)、F-actin蛋白表达升高(P<0.05),NF-κB p65入核明显增加。与LPS组相比,Apelin-13明显增加细胞活力(P<0.01),VE-cadherin蛋白表达增加(P<0.05)、F-actin蛋白表达下降(P<0.01),蛋白NF-κB p65入核下降明显。结论: Apelin-13可减轻LPS诱导的人脐静脉内皮细胞损伤及屏障受损,其机制可能与抑制炎症相关。

关键词: 内皮屏障, Apelin-13, 人脐静脉内皮细胞, 黏附连接, 炎症

Abstract: Objective: To investigate the effects of Apelin-13 on barrier function injury of human umbilical vein endothelial cells (HUVECs) induced by LPS. Methods: The HUVECs cultured in vitro were divided into 4 groups: Control group, LPS group, Apelin-13+LPS group, Apelin-13 group. HUVECs were treated by 5 μg/ml LPS for 24 h to replicate the model with endothelial barrier impaired. Apelin-13 at the concentration of 1 μmol/L was given 30 min before LPS treatment. The cell viabillity of HUVECs was measured by CCK-8 assay. Protein expressions of VE-cadherin and F-actin were measured by Western blot and immunofluorescence. Nuclear factor κB p65(NF-κB p65) was detected by immunofluorescence. Results: Compared with the control group, the cell viabillity of HUVECs and protein expression of VE-cadherin were decreased by LPS, but the protein expression of F-actin and activation of NF-κB p65 were increased by LPS. These effects were attenuated by Apelin-13 administration. Conclusion: Apelin-13 ameliorates LPS-induced barrier function injury of HUVECs, which may be related to the inhibition of inflammation.

Key words: endothelial barrier, Apelin-13, HUVECs, adherens junctions, inflammation

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