陆军军医大学学报 (Jun 2022)
Shared signaling pathway in pyroptosis and apoptosis in A549 cells induced by oxygen and glucose deprivation/recovery
Abstract
Objective To investigate the effects of oxygen and glucose deprivation/recovery (OGD/R) on apoptosis and pyroptosis of A549 cells, and to explore the possibility of the mutual regulation in apoptosis and pyroptosis. Methods A549 cells were resuscitated and cultured, randomly divided into control group, OGD/R group, OGD/R+pyroptosis inhibitor group (OGD/R+VX-765) and OGD/R+apoptosis inhibitor group (OGD/R+FMK). The apoptotic rate was detected by flow cytometry, the levels of pyroptosis markers IL-1β and IL-18 in the supernatant were measured by ELISA, and the expression of key molecules in pyroptosis and apoptosis at mRNA and protein levels were detected by RT-qPCR and Western blotting, respectively. Results The apoptotic rate was significantly increased in the OGD/R group than the control group (12.57±1.97 vs 3.67±0.64, P < 0.05), and the rate of the OGD/R+VX-765 group was decreased than that of the OGD/R group (5.97±1.28, P < 0.05). OGD/R resulted in obvious increased levels of IL-1β and IL-18 in the supernatant (17.35±0.44 vs 7.80±0.80 ng/L, 18.06±1.73 vs 7.99±0.11 ng/L, P < 0.05), but FMK treatment notably reversed the increases of 2 inflammatory factors (4.72±0.59, 10.24±1.42 ng/L, P < 0.05). OGD/R enhanced the mRNA and protein expression levels of key molecules of apoptosis and proptosis in A549 cells. While, VX-765 decreased the mRNA and protein levels of BID (1.12±0.18 vs 1.74±0.14, 1.01±0.10 vs 1.29±0.09, P < 0.05), and FMK reduced those of GSDME (0.94±0.05 vs 1.35±0.10, 0.52±0.03 vs 1.20±0.06, P < 0.05) when compared with the cells from the OGD/R group. Conclusion OGD/R can induce pyroptosis and apoptosis in A549 cells simultaneously, and there exists shared signaling pathway in pyroptosis and apoptosis. Pyroptosis and apoptosis inhibitors can simultaneously inhibit the 2 types of OGD/R-induced cell death.
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