Effect of Xinji Erkang on Human Umbilical Vein Endothelial Cells in Jury Induced by Angiotensin Ⅱ

CAI Guo-wei; CHENG Pan; HUANG Guang-yao; LIAN Feng-zhen; WANG Xiao-yun; HU Juan; ZHANG Yong-xue; GAO Shan

doi:10.13422/j.cnki.syfjx.20180929

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Effect of Xinji Erkang on Human Umbilical Vein Endothelial Cells in Jury Induced by Angiotensin Ⅱ

更新时间：2024-01-04

- Effect of Xinji Erkang on Human Umbilical Vein Endothelial Cells in Jury Induced by Angiotensin Ⅱ
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 9, Pages: 103-110(2018)
- 作者机构：
- 作者简介：
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- DOI：10.13422/j.cnki.syfjx.20180929
  CLC： R285
- Published：2018
- 稿件说明：
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CAI Guo-wei, CHENG Pan, HUANG Guang-yao, et al. Effect of Xinji Erkang on Human Umbilical Vein Endothelial Cells in Jury Induced by Angiotensin Ⅱ[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(9): 103-110.
DOI：

CAI Guo-wei, CHENG Pan, HUANG Guang-yao, et al. Effect of Xinji Erkang on Human Umbilical Vein Endothelial Cells in Jury Induced by Angiotensin Ⅱ[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(9): 103-110. DOI： 10.13422/j.cnki.syfjx.20180929.

摘要

目的：研究心肌尔康（XJEK）对血管紧张素Ⅱ（AngⅡ）诱导人脐静脉内皮细胞（HUVECs）损伤的作用，并探讨相关的作用机制。方法：体外培养HUVECs细胞，随机分为7组，分别为空白组，AngⅡ（1×10-5 mol · L-1）组，AngⅡ（1×10-5 mol · L-1）+心肌尔康不同质量浓度组（0.1，0.2，0.4，0.8，1.6 g · L-1），噻唑蓝（MTT）比色法检测HUVECs的活性；Griess法测定一氧化氮（NO）含量；流式细胞仪检测细胞内活性氧（ROS）的水平；钙成像仪测定细胞胞浆内游离钙离子（Ca2+）浓度；比色法及TBA法检测超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量；蛋白免疫印迹法（Western blot）检测内皮型一氧化氮合酶（eNOS）蛋白表达水平。结果：与空白组比较，AngⅡ组内皮细胞活力，NO释放，eNOS蛋白表达及SOD活力明显降低，MDA，ROS含量和内皮细胞胞浆内Ca2+浓度明显升高（P<0.05，P<0.01）；与AngⅡ组比较，心肌尔康可剂量依赖性提高内皮细胞活力，促进NO释放、增加eNOS蛋白的表达；升高SOD活力，抑制MDA，ROS含量和内皮细胞胞浆内Ca2+浓度的升高，各指标差异均具有显著性（P<0.05，P<0.01）。结论：心肌尔康对AngⅡ诱导的内皮损伤具有明显的保护作用，其可能机制为抑制细胞内Ca2+超载和降低氧化应激水平。

Abstract

Objective: To investigate the effect of Xinji Erkang (XJEK) on human umbilical vein endothelial cells (HUVECs) injury induced by angiotensin Ⅱ (AngⅡ). Method: HUVECs were cultured in vitro and randomly divided into 7 groups as follows:control group

AngⅡ (1×10-5 mol · L-1) group

and AngⅡ (1×10-5 mol · L-1)+XJEK groups (0.1

0.2

0.4

0.8

1.6 g · L-1). Thiazole blue (MTT) assay was used to examine HUVEC viability. The level of reactive oxygen species (ROS) and the intracellular free calcium concentration were measured by flow cytometry and Calcium Imager respectively. The content of nitric oxide (NO)

malondialdehyde (MDA) and superoxide dismutase (SOD) were detected by colorimetric and TBA analysis. Western blot was applied to determine the expression of endothelial nitric oxide synthase (eNOS) protein. Result: Compared with the control group

the Ang Ⅱ group showed significant reduction in endothelial cell vitality

NO release and eNOS protein expression

and significant increase in MDA and ROS content and Ca2+ concentration in endothelial cell cytoplasm (P<0.05

P<0.01). Compared with the AngⅡ group

XJEK could obviously improve endothelial dysfunction (ED) by promoting eNOS activities and enhancing NO in a dose-dependent manner. Moreover

XJEK could up-regulate SOD activity and down-regulate MDA content significantly. In addition

after treatment with XJEK

ROS level and intracellular Ca2+ concentration in endothelial cells were decreased compared with the AngⅡgroup (P<0.05). Conclusion: These results suggest that XJEK has a protective effect on AngⅡ-induced HUVECs injury

and the mechanism underlying may contribute to inhibiting intracellular Ca2+ overload and improving ED and ameliorating ROS.

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