Myeloid-derived growth factor improves pyroptosis in H9c2 cells induced by high glucose

Abstract

Objective To investigate whether myeloid-derived growth factor (Mydgf) can improve cardiomyocyte pyroptosis induced by hyperglycemia by regulating nucleotide-binding domain-like receptor protein 3 (NLRP3). Methods Cultured rat cardiomyocyte H9c2 cells were divided into 4 groups: normal group (Nor), high glucose group (HG, 50 mmol/L glucose for 24 h), Mydgf group (25 ng/mL recombinant Mydgf protein for 12 h, followed by 50 mmol/L glucose for 24 h), and necrosulfonamide group (NSA, 32 μmol/L necrosulfonamide for 0.5 h, followed by 50 mmol/L glucose for 24 h). Western blotting was used to detect the protein levels of cleaved Caspase-1, NLRP3, apoptosis-related speckle-like proteins (ACS) and gasdermin D (GSDMD). The secretions of IL-1β and IL-18 in cell culture medium were detected by ELISA. CCK-8 assay was used to detect the effect of Mydgf or NLRP3 on the viability of H9c2 cells. Lentiviral vector carrying NLRP3 and enhanced green fluorescent protein (EGFP) (NLRP3-EGFP) and empty viral vector (EGFP) were transfected into H9c2 cells respectively, and the changes of pyroptosis related indexes were detected by recovery experiments. Results Western blot results showed that the expression levels of pyroptosis related proteins, cleaved Caspase 1, ACS, NLRP3 and GSDMD were significantly higher in the HG group than the Nor group (P 0.05). In the recovery experiment, Western blot results showed that in the high-glucose environment, the expression level of NLRP3, ASC, and cleaved Caspase-1 were significantly increased (P < 0.05), and the secretions of IL-1β and IL-18 were also significantly enhanced in the Mydgf +NLRP3-EGFP group compared with the Mydgf group (P < 0.05). Conclusion Mydgf may ameliorate cardiomyocyte pyroptosis induced by hyperglycemia by inhibiting NLRP3.

Keywords

• myeloid-derived growth factor
• h9c2 cells
• pyroptosis
• nlrp3
• diabetic cardiomyopathy