Effect of estrogen-related receptor α on lipopolysaccharide-induced vascular endothelial apoptosis and tight junction protein degradation
10.3760/cma.j.issn.1671-0282.2021.11.006
- VernacularTitle:雌激素相关受体α对脂多糖诱导的内皮细胞凋亡及连接蛋白降解的影响
- Author:
Zhou PAN
1
;
Guang LI
;
Wei WU
;
Changyong WANG
;
Zhou LIU
;
Wenfang XIA
Author Information
1. 武汉大学人民医院重症医学科 430060
- Keywords:
Estrogen-related receptor α;
Acute lung injury;
Apoptosis;
Tight junction protein
- From:
Chinese Journal of Emergency Medicine
2021;30(11):1312-1317
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect of estrogen-related receptor α (ERRα) on lipopolysaccharide (LPS)-induced vascular endothelial cell apoptosis and tight junction protein degradation.Methods:RPMVECs transfected with shERRα were cultured in vitro and divided into four groups: Normal control group (Ctr group); shERRα knockdown group (shERRα group); normal cells + LPS treated group (LPS group): The cells in the six-well plates were cultured in serum-free medium for 12 h, and then treated with 20 μg/mL LPS for 12 h; and shERRα+LPS group: ERRα knockdown cells were treated as the LPS group. ROS fluorescence kit was used to detect the intracellular ROS levels . Apoptosis ratio was detected by TUNEL staining, AnnexinV-FITC and PI. Cell membrane ZO-1 expression was detected by cellular immunofluorescence, and the levels of apoptosis-related proteins Bcl-2, Bax, Smac, Cytochrome c, and tight junction protein ZO-1, as well as the expression of Occludin, JAM-A and E-Ca at molecular level were detected by Western blot.Results:Compared with the Ctr group and the shERRα group, the ROS level, apoptosis rate (TUNEL test: 16.44 ± 2.55; and flow cytometry test: 23.56 ± 2.22), the expression of pro-apoptotic proteins Bax, Smac and Cytochrome c were increased in the LPS group, while the expression of anti-apoptotic proteins Bcl-2 and tight junction protein were decreased. In the LPS group. Cellular immunofluorescence results showed that the ZO-1 was degraded in the cell membrane and the network structure was broken. Compared with the LPS group, inhibition of ERRα in the shERRα+LPS group increased cell damage.Conclusions:ERRα can negatively regulate the apoptosis and affect the function of pulmonary microvascular endothelial cells, thereby regulating sepsis-induced acute lung injury.
