Effects and mechanism of estrogen receptor β agonist on the migration and oxidative stress of human umbilical vein endothelial cell under high glucose condition
10.3760/cma.j.cn501120-20200720-00352
- VernacularTitle:雌激素受体β激动剂对高糖作用下人脐静脉内皮细胞迁移和氧化应激的影响及其机制
- Author:
Xiaoyu GUO
1
;
Xueqing ZHOU
;
Weiguo XIE
Author Information
1. 武汉大学同仁医院暨武汉市第三医院烧伤研究所 430060
- Keywords:
Estrogen receptor beta;
Endothelial cells;
Cell migration assays;
Oxidative stress;
High glucose
- From:
Chinese Journal of Burns
2021;37(9):869-874
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects and related mechanism of estrogen receptor β (ERβ) agonist on the migration and oxidative stress of human umbilial vein endothelial cells (HUVECs) under high glucose condition.Methods:The experimental research method was adopted. HUVECs were routinely cultured and passaged, and then cells of the logarithmic growth phase were collected for the subsequent experiments. The cells were divided into three groups according to the random number table, including normal control group (cultured with Roswell Park Memorial Institute 1640 cell culture medium (the same cell culture medium below) containing 5.5 mmol/L D-glucose), high glucose alone group (cultured with cell culture medium containing 25.0 mmol/L D-glucose alone), and high glucose+ERβ agonist diarylpropionitrile (DPN) group (cultured with cell culture medium containing 25.0 mmol/L D-glucose and 10 μmol/L DPN). Scratch test was conducted to detect the cell migration rate in the 3 groups at 24 h post scratching. Fluorescent probe method was used to detect the reactive oxygen species (ROS, denoted by red fluorescence intensity) of cells in the 3 groups on 5 d post culture. Western blotting was used to detect the protein expression levels of vascular endothelial growth factor (VEGF) and superoxide dismutase 2 (SOD2) of cells in the 3 groups on 5 d post culture. In the above-mentioned experiments, cells were grouped and cultured correspondingly as before, the number of samples in each group was 5. Data were statistically analyzed with one-way analysis of variance and least significant difference t test. Results:At 24 h post scratching, the cell migration rate in high glucose alone group was (36±5)%, which was significantly lower than (76±4)% of normal control group and (65±5)% of high glucose+DPN group ( t=14.511, 9.603, P<0.01), and the cell migration rate in high glucose+DPN group was significantly lower than that in normal control group ( t=3.943, P<0.01). On 5 d post culture, the level of ROS of cells in high glucose alone group (1.81±0.12) was significantly increased compared with normal control group and high glucose+DPN group (1.00±0.14, 0.91±0.15, t=9.679, 10.549, P<0.01), while the level of ROS of cells in normal control group and high glucose+DPN group were close ( t=1.031, P>0.05). On 5 d post culture, the protein expression levels of VEGF and SOD2 of cells in high glucose alone group were significantly lower than the levels of normal control group ( t=14.175, 13.787, P<0.01) and high glucose+DPN group ( t=6.321, 17.750, P<0.01). The protein expression level of VEGF of cells in high glucose+DPN group was significantly lower than the level of normal control group ( t=7.206, P<0.05), while the protein expression level of SOD2 of cells in high glucose+DPN group was significantly higher than the level of normal control group ( t=2.890, P<0.05). Conclusions:The activation of ERβ can improve the inhibition of HUVECs migration induced by high glucose and alleviate oxidative stress injury induced by high glucose, which may be achieved by promoting the expression of VEGF and SOD2.
