GSDMD-miR-223-NLRP3 axis involved in B(a)P-induced inflammatory injury of alveolar epithelial cells

• Published in: Ecotoxicology and environmental safety 2022-03, Vol.232, p.113286-113286, Article 113286
• Main Authors: Kong, Xiangbing, Gao, Min, Liu, Yitong, Zhang, Peng, Li, Mengyuan, Ma, Pengwei, Shang, Pingping, Wang, Wei, Liu, Hong, Zhang, Qiao, Feng, Feifei
• Format: Article
• Language: English
• Subjects: Alveolar Epithelial Cells - metabolism; Benzo(a)pyrene; Cell Line, Tumor; GSDMD; Humans; Inflammasomes - metabolism; Interleukin-1beta - metabolism; MicroRNAs - genetics; MicroRNAs - metabolism; miR-223; NLR Family, Pyrin Domain-Containing 3 Protein - genetics; NLR Family, Pyrin Domain-Containing 3 Protein - metabolism; NLRP3 inflammasome; Pyroptosis
• ISSN: 0147-6513; 1090-2414
• DOI: 10.1016/j.ecoenv.2022.113286

Benzo(a)pyrene [B(a)P], a ubiquitous environmental pollutant, causes lung inflammatory damage. Pyroptosis，a new inflammation-dependent programmed cell death, happened when pyroptosis-related GSDMD is activated mediated by NLRP3 inflammasome. microRNA-223 (miRNA-223) is involved in inflammatory diseases by regulating NLRP3. However, whether GSDMD regulate NLRP3 inflammasome through miR-223 in B(a)P induced lung inflammatory injury remain unknown. In this study, alveolar epithelial cells (A549) were stimulated with 0, 2, 4, 8 μM B(a)P for 12 h or 24 h. The inflammatory injury and pyroptosis were determined. And the activation of NLRP3 inflammasomes and the level of miRNA-223 were detected. Then, the change of inflammatory injury and activation of NLRP3 inflammasomes in B(a)P-induced A549 cells were detected after inhibiting of GSDMD or miR-223 using siRNA-GSDMD (siGSDMD) or miR-223 inhibitor, respectively. Our results indicated that after B(a)P exposure, TNF-α and IL-6 in the supernatant were increased. Transmission electron microscope (TEM) results showed that A549 cells were obviously swollen and the cell membrane ruptured. Hoechest33342/PI staining showed that pyroptosis occurred. NLRP3, IL-1β, IL-18, GSDMD, GSDMD-N, pro caspase-1 and cleaved caspase-1 were significantly increased. Additionally, after transfecting with siGSDMD in B(a)P-induced A549 cells, the expression level of miR-223 was significantly increased. But IL-6 and TNF-α in the supernatant, the expression of NLRP3, IL-1β, IL-18 and cleaved caspase-1 protein were also decreased. And after inhibiting miR-223 in B(a)P-induced A549 cells, the expression of TNF-α and IL-6 in the supernatant, the protein expression of NLRP3, IL-1β, IL-18 and cleaved caspase-1 were increased. In conclusion, GSDMD may regulate NLRP3 inflammasome through miR-223, which is involved in B(a)P induced inflammatory damage in A549 cells. [Display omitted] •Pyroptosis occurred simultaneously when B(a)P-induced inflammatory injury in alveolar epithelial A549 cells.•NLRP3 and miRNA-223 were inhibited when knockout of GSDMD in B(a)P-induced A549 cells.•NLRP3 were increased when inhibiting miR-223 in B(a)P-induced A549 cells.•GSDMD may regulate NLRP3 inflammasome through miR-223 in B(a)P induced inflammatory damage in A549 cells.