Glucocorticoid induces GSDMD-dependent pyrolysis in PC12 cells via endoplasmic reticulum stress

Abstract

Objective The present study explored whether pyroptosis is involved in the injury process of PC12 cells induced by glucocorticoid (GC) and the regulatory relationship between endoplasmic reticulum stress (ERS) and pyrolysis. Methods LDH leakage of PC12 cells was detected by LDH assay. The number of dead cells was detected by SYTOX green nucleic acid staining. The levels of IL-1β and IL-18 in the supernatants was detected by ELSIA assay. The expression levels of glucose regulated protein 78 (GRP78), cleaved gasdermin D-NT (cleaved-GSDMD-NT), NLR-pyrin domain-containing 3 (NLRP3) and cleaved-caspase-1 were observed by immunofluorescence staining and western blot. Results The LDH assay revealed that GC exposure significantly increased the release of LDH. The results of SYTOX green acid staining showed that GC exposure significantly increased the number of SYTOX green acid-positive cells. The ELSIA assay revealed that GC exposure significantly increased the levels of IL-1β and IL-18 in the supernatants. The results of immunofluorescence staining and western blot showed that GC exposure significantly increased the expression of GRP78, cleaved-GSDMD-NT, NLRP3 and cleaved caspase-1. Treatment with the ERS inhibitor tauroursodeoxycholate (TUDCA) and siRNA GSDMD attenuated related damage and downregulated the expression of the abovementioned proteins. Conclusion The present study clearly demonstrated that GC exposure can induce GSDMD-dependent pyrolysis, and ERS is involved in the above damage process.

Inflammasome Activity in the Skeletal Muscle and Heart of Rodent Models for Duchenne Muscular Dystrophy

“…In line with the previous findings, GSDMD showed mild elevation in the skeletal muscle in our study, suggesting that pyroptosis may contribute to muscle loss in DMD. Interestingly, GSDMD-mediated pyroptosis is associated with glucocorticoidinduced muscular atrophy via various mechanisms [29,30], which is in clear contrast to the clinical observation that glucocorticoids are beneficial in DMD patients. In fact, glucocorticoid-mediated protection was also reported along with other drugs [31].…”

Gasdermins in sepsis

“…GSDMs-mediated pathways are regulated by various molecules, such as miRNAs, oxidative stress, chemokines (83)(84)(85)(86). Activated GSDMs proteins bound to the plasma membrane and formed pores (87), mediating cell death or inflammatory factors release, functional proteins, etc.…”