Effect of Cryptotanshinone on Hyperglycemic-induced Pyroptosis of Cardiomyocytes by Regulating TLR4/NF-κB/NLRP3 Signaling Pathway
Objective:To investigate the effect of cryptotanshinone regulating Toll-like receptor 4/nuclear transcription factor-κB/NOD-like receptor protein 3(TLR4/NF-κB/NLRP3)signaling pathway on hyperglycemic-induced pyroptosis of cardiomyocytes.Methods:H9c2 cells were divided into control group(normal glucose 5.5 mmol/L),high glucose group(glucose 33 mmol/L),2.5 μmol/L cryptotanshinone group(glucose 33 mmol/L+2.5 μmol/L cryptotanshinone)and 5 μmol/L cryptotanshinone group(glucose 33 mmol/L+5 μmol/L cryptotanshinone)and lipopolysaccharide(LPS)group(33 mmol/L+5 μmol/L cryptotanshinone+1 μg/mL TLR4/NF-κB/NLRP3 signaling pathway activator LPS).The cells of each group were cultured in the corresponding medium for 48 h.The cell survival rate was detected by MTT method.The level of reactive oxygen species(ROS)was detected by 2'7'-diacetyldichlorofluorescein(DCFH-DA)method.Cysteine aspartase 1 inhibitor(FLICA)-cysteine aspartase 1(Caspase-1)/propyl iodide(PI)double staining method was used to detect the pyroptosis of cells.The levels of interleukin-6(IL-6)and interleukin-1β(IL-1β)were detected by enzyme-linked immunosorbent assay(ELISA).The N-terminal fragment of GSDMD protein(GSDMD-N),TLR4,NLRP3,phosphorylated NF-κB p65(p-NF-κB p65),NF-κB p65,Cleaved Caspase-1,and Caspase-1 protein expression were detected by Western Blot.Results:Compared with the control group,the survival rate of H9c2 cells in the high glucose group decreased(P＜0.05),while the level of ROS,proportion of pyrodead cells,levels of IL-1β and IL-18,the expressions of TLR4,NLRP3,P-NF-κB p65/NF-κB p65,GSDMD-N,Cleaved Caspase-1/Caspase-1 protein increased(P＜0.05).Compared with the high glucose group,the survival rate of H9c2 cells in the 2.5 μmol/L cryptotanshinone group and the 5 μmol/L cryptotanshinone group increased(P＜0.05),while the level of ROS,proportion of pyrodead cells,levels of IL-1βand IL-18,the expressions of TLR4,NLRP3,P-NF-κB p65/NF-κB p65,GSDMD-N,Cleaved Caspase-1/Caspase-1 protein decreased(P＜0.05).TLR4/NF-κB/NLRP3 signaling pathway activator LPS showed reversing the inhibitory effect of cryptotanshinone on hyperglucose induced cell proliferation and pyroptosis.Conclusion:Cryptotanshinone could inhibit TLR4/NF-κB/NLRP3 signaling pathway to reduce hyperglycemic-induced myocardial pyroptosis.
cardiomyocytescryptotanshinoneToll-like receptor 4/nuclear transcription factor κB/NOD-like receptor protein 3 signaling pathwayhyperglycemicpyroptosisexperimental study
NETL
NSTL
万方数据
