Effects of Oxygen-Glucose Deprivation/Recovery on Pyroptosis of A549 Alveolar Epithelial Cells

WANG Xiaoyan, XIAO Zongyi, YI Han, AN Yuxuan, SONG Juan, CHEN Fei, WANG Shouyong*

(Department of Anesthesiology, Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing 400014, China)

Abstract:

To further explore the mechanisms of cardiopulmonary bypass-related acute lung injury, an OGD/R (oxygen-glucose deprivation/recovery) model using A549 alveolar epithelial cell has been established to simulate the ischemia-reperfusion process of alveolar epithelial cell during clinical cardiopulmonary bypass. The pyroptosis of A549 cell caused by OGD/R was observed. The mRNA and protein expression levels of pyroptosis related gene Caspase1, ASC, NLRP3, GSDMD were detected by RT-qPCR and Western blot. Then the effects of Caspase1 inhibitor VX-765 on Caspase1 and GSDMD expression, the A549 cell survival rate, as well as the concentrations of IL-1β and IL-18 in cell culture supernatant were measured, respectively. The results showed that OGD/R induced an significant increase in Caspase1 activity (P<0.05), and the mRNA and protein expression levels of pyroptosis-related gene Caspase1, ASC, NLRP3, GSDMD were up-regulated significantly (P<0.05). When pretreatment with the Caspase1 inhibitor VX-765, the Caspase1 and GSDMD expression levels, the concentration of IL-1β and IL-18 in cell culture supernatant were decreased, and the A549 cell survival rate after OGD/R was restored. This study suggests that OGD/R induces A549 alveolar epithelial cell pyroptosis, and inhibition of the occurrence of pyroptosis has a protective effect on OGD/R cell model, which may be involved in the mechanisms of cardiopulmonary bypass-related lung injury.