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Effect of estrogen-related receptor α on lipopolysaccharide-induced vascular endothelial apoptosis and tight junction protein degradation / 中华急诊医学杂志
Chinese Journal of Emergency Medicine ; (12): 1312-1317, 2021.
Article in Chinese | WPRIM | ID: wpr-907769
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.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Emergency Medicine Year: 2021 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Emergency Medicine Year: 2021 Type: Article