Study On The Mechanism Of Dapagliflozin On Palmitic Acid-induced Pyroptosis Of Glomerular Podocyte

Objective: To investigate the effect and mechanism of dapagliflozin on pyroptosis in MPC-5 glomerular podocytes induced by palmitic acid high fat,and to provide a new theoretical basis for analyzing the protective effect of diabetic nephropathy.Methods: Cells were induced with different concentrations of PA to establish a high-fat model.Western blot analysis was performed to assess the changes in levels of pyroptosis-related proteins,including NLRP3,GSDMD,Caspase-1,IL-18,IL-1β,and ASC.The cells were divided into Con,Con+Dapa,PA,and PA+Dapa groups.After treatment with Dapagliflozin,Western blot analysis was conducted to determine the changes in levels of pyroptosis-related proteins(NLRP3,Caspase-1,IL-1β,ASC)and m RNA.Furthermore,the cells were divided into Con,Con+HO-1,PA,and PA+HO-1groups.The MPC5 cells were transfected with the overexpression plasmid p CDNA3.1-ho-1:t2a:egf and subsequently treated.Western blot analysis was performed to evaluate the changes in related proteins such as NLRP3,GSDMD,Caspase-1,HO-1,and m RNA levels.Results: We established a cellular high-fat model induced by PA and observed an upregulation of protein expression for NLRP3,Caspase-1,IL-1β,ASC,and other pyroptosis-related proteins under PA induction,reaching a peak at a PA concentration of 0.2 m M.When treated with Dapagliflozin,significant downregulation in the expression levels of NLRP3,GSDMD,Caspase-1,IL-18,IL-1β,ASC,and other pyroptosis-related proteins,as well as m RNA levels,was observed compared to the PA group.Conversely,the protein expression and m RNA levels of HO-1 were significantly upregulated.Following transfection with HO-1 overexpression,Western blot analysis showed an upregulation in the expression of pyroptosis-related factors such as NLRP3,Caspase-1,IL-1β,ASC,while HO-1 protein expression was downregulated.q RT-PCR results were consistent with these findings.In comparison to the PA group,there was a significant downregulation in the expression levels of NLRP3,GSDMD,Caspase-1,IL-18,IL-1β,ASC,and other pyroptosis-related proteins,as well as m RNA levels,while HO-1 protein expression and m RNA levels were significantly upregulated.Conclusion: PA induction resulted in cell pyroptosis,while treatment with Dapagliflozin suppressed the expression of pyroptosis-related factors,thereby alleviating cell pyroptosis.Overexpression of HO-1 also inhibited the expression of pyroptosis-related factors in PA-induced MCP5 cells,thereby attenuating pyroptosis.Dapagliflozin may potentially exert its effects on cell pyroptosis by mediating the expression of HO-1.