Study on mechanism of AGEs mediating inflammation of cardiomyocytes through TLR4/MD2 pathway
Objective To explore the mechanisms by which advanced glycation end products(AGEs)mediate inflamma-tion of cardiomyocytes through the toll-like receptor 4(TLR4)/myeloid differentiation factor 2(MD2)pathway.Methods Intervention for 12,24,48,60,72 h was performed using different concentrations of AGEs bovine serum albumin(0,100,200,400,800 μg/mL)to identify the optimal AGEs concentration and intervention time.Subsequently,H9c2 cells were divided into four groups:control group,AGEs group,MD-2 inhibitor group,and AGEs+MD2 inhibitor group.The cell viability of H9c2 cells in each group was assessed using the CCK-8 assay.LDH levels in the culture medium of H9c2 cells were measured u-sing a reagent kit.The concentrations of AGEs-MD2 in cardiomyocytes were measured by ELISA.The expression levels of MyD88 protein downstream of TLR4 in cardiomyocytes were detected by Western blot.The mRNA expression levels of IL-6 and TNF-α of inflammatory markers in cardiomyocytes were determined by real time PCR.Results Compared with the control group,the AGEs group showed a decrease in cell viability,an increase in LDH release,optical density at 450 nm for AGEs-MD2,relative protein expression of MyD88,and an increase in relative mRNA expression of IL-6 and TNF-α,and all the differ-ences were statistically significant(P＜0.05).Compared with the control group,the MD-2 inhibitor group showed a decrease in cell viability and an increase in LDH release,and all the differences were statistically significant(P＜0.05).Compared with the AGEs group,the AGEs+MD2 inhibitor group showed a decrease in optical density at 450 nm for AGEs-MD2,a decrease in rel-ative protein expression of MyD88,and a decrease in relative mRNA expression of IL-6 and TNF-α,and all the differences were statistically significant(P＜0.05).Conclusion AGEs can activate cardimyocyte inflammation which is mediated by the inflam-matory cytokines of MyD88 through the TLR4/MD2 pathway,which may be one of the mechanisms of cardiac remodeling caused by the long-term accumulation of AGEs in the body.
advanced glycation end productsadvanced glycation end product receptortoll-like receptor 4myeloid differ-entiation receptor 2
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