Effect of Andrographolide on High Glucose Induced Pyroptosis of Myocardial Cells by Regulating the AMPK/NLRP3 Signaling Pathway
SUN Xue
LIU Huimin
RUAN Hualing
ZHANG Bingfei
Abstract：Objective:To investigate the effect of andrographolide(AD)on high glucose(HG)induced pyroptosis of myocardial cells by regulating the adenosine monophosphate-activated protein kinase(AMPK)/Nod-like receptor protein 3(NLRP3)signaling pathway.Methods:Logarithmic phase H9c2 cells were divided into control group,HG group(50 mmol/L HG),HG+AD-L group(30 mg/L AD+50 mmol/L HG),HG+AD-M group(40 mg/L AD+50 mmol/L HG),HG+AD-H group(50 mg/L AD+50 mmol/L HG),and HG+AD-H+AMPK inhibitor Compound C(CC)group(50 mg/L AD+50 mmol/L HG+10 μmol/L CC).CCK-8 method and Hoechst 33342/propidium iodide(PI)staining were used to determine the proliferation activity and cell pyroptosis of H9c2 cells.Enzyme-linked immunosorbent assay(ELISA)method was applied to determine the levels of lactate dehydrogenase(LDH)and inflammatory factors.Reverse transcription polymerase chain reaction(qRT-PCR)was applied to measure the mRNA expression of cell pyroptosis factor.Western Blot was applied to determine the expression of AMPK/NLRP3 pathway proteins and pyroptosis proteins.Results:10-60 mg/L of AD was able to enhance the HG-induced proliferation activity of H9c2 cells,and after intervention with 50 mg/L of AD,the proliferation activity of H9c2 cells was the highest.Therefore,30,40,and 50 mg/L of AD were selected for subsequent experiments.Compared with the control group,the OD570 value and p-AMPK/AMPK in the HG group decreased,while the cell pyroptosis rate,LDH,levels of inflammatory factors,mRNA and protein expression of pyroptosis factors increased(P＜0.05).Compared with the HG group,the OD570 value and p-AMPK/AMPK in the HG+AD-L group,HG+AD-M group,and HG+AD-H group all increased,while the remaining indicators mentioned above decreased(P＜0.05).Compared with the HG+AD-H group,the OD570 value and p-AMPK/AMPK in the HG+AD-H+CC group decreased,while the remaining indicators above all increased(P＜0.05).Conclusion:AD can alleviate HG-induced inflammatory damage to H9c2 cells,inhibit cell pyroptosis,and this effect may be related to regulating the AMPK/NLRP3 signaling pathway.
Keywords：cell pyroptosismyocardial cellshigh glucoseandrographolideAMP-activated protein kinase/Nod-like receptor thermal protein domain associated protein 3experiment research
Publication Date：2025-08-25
Online Publishing Date：2025-09-10(First online date of this platform, not the publication date of the document)
Pages：6( 2468-2473 )