Protective Effect of Total Flavonoids from Puerariae Lobatae Radix on Oxidative Stress-induced Injury of Human Umbilical Vein Endothelial Cells and Its Mechanism

SU Cheng-fu; GONG Man; HAN Yong-guang; WEI Yu; LI Yan-ling; ZHAO Yi-ying; SUN Yi-jie; LIU Ya-min

doi:10.13422/j.cnki.syfjx.20182335

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Protective Effect of Total Flavonoids from Puerariae Lobatae Radix on Oxidative Stress-induced Injury of Human Umbilical Vein Endothelial Cells and Its Mechanism

更新时间：2024-09-23

- Protective Effect of Total Flavonoids from Puerariae Lobatae Radix on Oxidative Stress-induced Injury of Human Umbilical Vein Endothelial Cells and Its Mechanism
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 23, Pages: 159-164(2018)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.20182335
  CLC： R285
- Published：2018
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SU Cheng-fu, GONG Man, HAN Yong-guang, et al. Protective Effect of Total Flavonoids from Puerariae Lobatae Radix on Oxidative Stress-induced Injury of Human Umbilical Vein Endothelial Cells and Its Mechanism[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(23): 159-164.
DOI：

SU Cheng-fu, GONG Man, HAN Yong-guang, et al. Protective Effect of Total Flavonoids from Puerariae Lobatae Radix on Oxidative Stress-induced Injury of Human Umbilical Vein Endothelial Cells and Its Mechanism[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(23): 159-164. DOI： 10.13422/j.cnki.syfjx.20182335.

摘要

目的：研究葛根总黄酮对H2O2诱导人脐静脉内皮细胞（HUVECs）氧化应激损伤的影响，并探讨其相关的作用机制。方法：以HUVECs为研究对象，实验分为空白组、模型组、阳性药组（10 μmol·L-1依达拉奉）和葛根总黄酮10，20，40，80，160 mg·L-1组；采用噻唑蓝（MTT）比色法检测葛根总黄酮对HUVECs的保护作用，采用分光光度法测定细胞培养上清液中细胞内超氧化物歧化酶（SOD），丙二醛（MDA）及NO含量；酶联免疫吸附测定（ELISA）检测细胞上清液中内皮素-1（ET-1）含量；采用二氯荧光乙酰乙酸盐（DCFH-DA）法检测活性氧（ROS）水平；蛋白免疫印迹法（Western blot）检测内皮型一氧化氮合酶（eNOS）和血管细胞黏附分子-1（VCAM-1）的表达。结果：与正常组比较，模型组细胞存活率显著降低（P<0.01），SOD活性，NO水平和eNOS蛋白表达显著降低（P<0.01），MDA，ET-1水平，细胞ROS水平和VCAM-1蛋白表达显著升高（P<0.01）；与模型组比较，葛根总黄酮（40，80，160 mg·L-1）可以显著提高氧化应激损伤的HUVECs的存活率（P<0.05，P<0.01），葛根总黄酮（20，40，80 mg·L-1）可显著增加SOD活性，升高NO水平，显著降低MDA，ET-1水平，显著降低细胞ROS水平，显著升高eNOS，降低VCAM-1的表达（P<0.05，P<0.01）。结论：葛根总黄酮对H2O2诱导HUVECs氧化应激损伤有保护作用，其作用机制可能与调节SOD，MDA，ET-1，NO氧化应激相关因子，调控内皮细胞功能相关蛋白eNOS，VCAM-1的表达有关。

Abstract

Objective:To study the effect of total flavonoids from Puerariae Lobatae Radix on oxidative stress injury induced by H2O2 in human umbilical vein endothelial cells (HUVECs). Method:With HUVECs as the study object

the experiment was divided into blank group

model group

positive drug group (10 μmol·L-1

Edaravone) and total flavone from Pueraria Lobatae Radix groups (10

160 mg·L-1). Methyl thiazolyl tetrazolium(MTT) assay was used to detect the protective effect of total flavonoids of Puerariae Lobatae Radix on HUVECs. The contents of superoxide dismutase (SOD)

malondialdehyde (MDA) and nitric oxide (NO) were detected by spectrophotometry

and the content of endothelin-1(ET-1) in the cell supernatant was detected by enzyme linked immunosorbent assay (ELISA). The level of reactive oxygen(ROS) was detected by DCFH-DA

and Western blot was used to detect the expressions of endothelial nitric oxide synthase (eNOS) and vascular cell adhesion molecule-1 (VCAM-1). Result:Compared with normal group

model group showed a significantly lower cell survival rate

activity of SOD

level of NO and eROS protein expression (P<0.01)

and a significantly higher MDA

ET-1 level

cell ROS level and VCAM-1 protein expression (P<0.01). Compared with model group

total flavone from Pueraria Lobatae Radix (40

160 mg·L-1) could significantly increase the survival rate of HUVECs with oxidative stress injury (P<0.05

P<0.01)

and total flavone from Pueraria Lobatae Radix (20

80 mg·L-1) could significantly increase NO level and eNOS

and significantly decrease MDA and ET-1 levels

cell ROS level and VCAM-1 expression (P<0.05

P<0.01). Conclusion:Total flavonoids from Puerariae Lobatae Radix can protect human umbilical vein endothelial cells from oxidative stress injury induced by H2O2.The mechanism may be related to the regulation of oxidative stress-related factors of SOD

MDA

ET-1 and NO

and the expressions of endothelial cell function-related proteins eNOS and VCAM-1.

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