基于ACE2-Ang(1-7)-Mas轴探讨定心方三号方改善人脐静脉内皮细胞功能的机制研究  

Mechanism of Dingxin Recipe Ⅲ on Improving Umbilical Vein Endothelial Cell Function by Regulating ACE2-Ang(1-7)-Mas Axis

作　　者：刘晓瑜[1] 程赛博[1] 徐煜凌 何培坤 顾民华 周凤华[1] 贾钰华[1] LIU Xiaoyu;CHENG Saibo;XU Yuling;HE Peikun;GU Minhua;ZHOU Fenghua;JIA Yuhua(College of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515 Guangdong, China)

机构地区：[1]南方医科大学中医药学院,广东广州510515

出　　处：《中药新药与临床药理》2019年第3期301-307,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金(81373574);广州市科技计划产学研协同创新重大专项项目(201704020042);广东省自然科学基金项目(2014A030313354);广东省省级科技计划项目(2016A020226002);广东省名优中成药二次开发项目(20174010)

摘　　要：目的探讨定心方三号方(Dingxin RecipeⅢ,DXRⅢ)是否能通过调节ACE2-Ang(1-7)-Mas轴对抗血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)引起的人脐静脉内皮细胞(Human umbilical vein endothelial cells,HUVECs)损伤。方法制备DXRⅢ含药血清,HUVEC细胞分为5组,除外对照组,其余组均采用10 mol·L-1的AngⅡ刺激建立HUVECs损伤模型,造模后,分别予以空白血清、5%DXRⅢ含药血清、10%DXRⅢ含药血清以及20%DXRⅢ含药血清进行干预。BCECF-AM荧光染色观察HUVECs的黏附性改变,Western Blot法检测血管紧张素转换酶2(angiotensin-coverting enzyme, ACE2)、 Mas、细胞间黏附分子1(intercellular cell adhesion molecule-1,ICAM-1)、血管细胞黏附分子(vascular cell adhesion protein 1,VCAM-1)和p-p38蛋白的表达,ELISA法检测细胞上清中IL-6和血管紧张素1-7[angiotensin1-7,Ang(1-7)]水平。结果单核细胞黏附实验结果显示黏附细胞数量随着DXRⅢ含药血清浓度增加而减少(P<0.01);Western Blot结果显示,DXRⅢ中、高剂量组可促进ACE2和Mas蛋白的表达(P<0.01),抑制ICAM-1、VCAM-1以及p-p38蛋白的表达(P<0.05);ELISA结果显示,DXRⅢ中、高剂量组可抑制IL-6释放(P<0.05),增加Ang(1-7)水平(P<0.01)。结论DXRⅢ可能通过调控ACE2-Ang(1-7)-Mas轴拮抗AngⅡ诱导的HUVECs损伤后p-p38磷酸化,进而抑制ICAM-1、VCAM-1和IL-6的表达,从而起到保护内皮细胞功能的作用。Objective To investigate whether Dingxin Ⅲ Recipe can protect angiotensin Ⅱ induced human umbilicalvein endothelial cells injury by regulating ACE2-Ang(1-7)-Mas axis. Methods HUVECs were divided into 5 groups,the DXR Ⅲ drug-containing serum was prepared. Except for the control group,the rest of the groups werestimulated with 10 μmol·L^-1 AngII to establish HUVECs injury model. After modeling,cells in each group weretreated with blank serum,5%,10% and 20% DXR Ⅲ drug-containing serum. The change of adhesion of HUVECswas observed by BCECF-AM fluorescence staining. Expressions of ACE2,Mas,ICAM-1,VCAM-1 and p-p38 proteins were detected by Western Blot. The levels of IL-6 and Ang(1-7) in cell supernatants were measured byELISA assay. Results Compare with the model group, the number of adherent cells decreased with increasingconcentration of DXR Ⅲ drug-containing serum(P < 0.01). ACE2 and Mas expression in 10% and 20% DXR Ⅲ groups were significantly enhanced(P < 0.01) while the expression of ICAM-1, VCAM-1 and p-p38 weresignificantly reduced(P < 0.05). Furthermore,the levels of IL-6 in 10% and 20% DXR Ⅲ groups were decreased(P < 0.05) while the levels of Ang(1-7) were increased(P < 0.01). Conclusion DXR Ⅲ may inhibitphosphorylation of p-p38,decrease the expression of ICAM-1,VCAM-1 and IL-6 by regulating ACE2-Ang(1-7)-Mas axis in HUVECs to protect the function of endothelial cells.

关 键 词：定心方三号方 ACE2 Ang(1-7) MAS 内皮细胞 

分 类 号：R285.5[医药卫生—中药学]