The Effect Of NLRP3 On The Oxymatrine-Inhibiting INS-1 Cells Pyroptosis

Objective:To explore the possible mechanism of the inhibition of pyroptosis in islet cells by oxymatrine,the effects of oxymatrine on the expression of pyroptosis related proteins were investigated by applying an inhibitor of Nod-like receptor family pyrin domain containing 3(NLRP3)in rat insulinoma cell(INS-1).The results provide a theoretical basis for the better application of oxymatrine in the treatment of diabetes.Methods:INS-1 cells were induced pyroptosis by glucose and palmitic acid sodium.The experiments were divided into control group,high-glucose + high-fat group(HG + PA),high-glucose + high-fat + oxymatrine group,high-glucose + high-fat + oxymatrine +NLRP3 inhibitor(MCC950)group and high-glucose + high-fat + MCC950 group.The effect of oxymatrine on the activity of INS-1 cells was studied by CCK-8 assay.The activity of Caspase-1 was tested by Caspase-1 activity assay kit.The protein expression levels of NLRP3 and GSDMD were detected by Western blotting assay.Results:In CCK-8 experiment,the activity of INS-1 cells in HG + PA group decreased(**P<0.01 vs control group).Compared with the HG + PA group,the activity of INS-1 cells in the HG + PA + oxymatrine group,the HG + PA + oxymatrine + MCC950 group and the HG + PA + MCC950 group increased(**P<0.01 vs HG + PA group).Compared with the HG + PA + MCC950 group,there was no significant change in cell activity in the HG + PA+ oxymatrine + MCC950 group(P > 0.05 vs HG + PA + MCC950 group).Compared with the HG + PA + oxymatrine group,there was no significant change in cell activity in the HG+ PA + oxymatrine + MCC950 group(P > 0.05 vs HG + PA + oxymatrine group).In the Caspase-1 activity assay,Caspase-1 activity of INS-1 cells increased in the HG + PA group(**P<0.01 vs control group).Compared with the HG + PA,Caspase-1 activity of INS-1 cells decreased in the HG + PA + oxymatrine group,the HG + PA + oxymatrine + MCC950 group and the HG + PA +MCC950 group(**P<0.01 vs HG + PA group).Compared with the HG+ PA + MCC950 group,Caspase-1 activity of INS-1 cells in the HG + PA + oxymatrine +MCC950 group was not significantly changed(P > 0.05 vs HG + PA + MCC950 group).Compared with the HG + PA + oxymatrine group,Caspase-1 activity of INS-1 cells in the HG + PA + oxymatrine + MCC950 group was not significantly changed(P > 0.05 vs HG +PA + oxymatrine group).In Western blotting assay,the protein levels of NLRP3 and GSDMD-N in the HG + PA group were increased(**P<0.01 vs control group),GSDMDtotal protein had no significant change(P > 0.05 vs control group).Compared with the HG+ PA group,the protein expressions of NLRP3 and GSDMD-N in the HG + PA + oxymatrine group,the HG + PA + oxymatrine + MCC950 group and the HG + PA + MCC950 group decreased(**P<0.01 vs HG + PA group).Compared with the HG + PA + MCC950 group,the protein levels of NLRP3 and GSDMD-N in the HG + PA + oxymatrine + MCC950 group were not significantly changed(P> 0.05 vs HG + PA + MCC950 group).Compared with the HG + PA + oxymatrine group,the protein levels of NLRP3 and GSDMD-N in the HG + PA+ oxymatrine + MCC950 group were not significantly changed(P> 0.05 vs HG + PA +oxymatrine group).Conclusion:Oxymatrine inhibits the pyroptosis of INS-1 cells through NLRP3 in high glucose and high fat.This conclusion is helpful to deepen the theory of oxymatrine-regulated islet cell function and provide theoretical basis for its treatment of diabetes.