Effects of Glycyrrhizin Modulating HMGB1/RAGE Signaling Pathway on Apoptosis and Autophagy of Cardiomyocytes Induced by High Glucose
Objective:To investigate the effect of glycyrrhizin(Gly)on apoptosis and autophagy induced by high glucose in cardiomyocytes by regulating the high mobility group box protein(HMGB1)/receptor for advanced glycation end products(RAGE)signaling pathway.Methods:Rat cardiomyocytes H9c2 were cultured in vitro and divided into control(Ctrl)group,high glucose group,high glucose+Gly group,high glucose+pcDNA3.1 empty vector group,high-glucose+pcDNA3.1-HMGB1 group,high glucose+Gly+pcDNA3.1 empty vector group,and high glucose+Gly+pcDNA3.1-HMGB1 group.Cell viability was detected by cell counfing kit-8(CCK-8)assay.Apoptosis rate was detected by Annexin V-FITC/propyl iodide(PI)method.Autophagy was observed by transmission electron microscopy.The expression of B-cell lymphoma/leukemia gene-2-related X protein gene(Bax),microtubule-related protein 1 light chain 3(LC3)and Beclin-1 apoptosis and autophagy-related protein were detected by Western Blot.The levels of interleukin-1β(IL-1β)and tumor necrosis factor α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).2,7-dichlorofluorescein diacetate(DCFH-DA)fluorescence probe method was performed to detect reactive oxygen species(ROS).Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was performed to detect the mRNA expression levels of HMGB1 and RAGE.Results:Compared with Ctrl group,H9c2 cell viability inhigh glucose group decreased,apoptosis rate,autophagosome number and protein expression of Bax,LC3 and Beclin-1 increased,IL-1β,TNF-α,ROS levels and mRNA expression of HMGB1 and RAGE increased,with statistical significance(P＜0.05).Compared with the high glucose group,the H9c2 cell viability was increased in the high glucose+Gly group,the apoptosis rate,the number of autophagosomes and the expression of Bax,LC3 and Beclin-1 protein decreased,and the levels of IL-1β,TNF-α and ROS and the mRNA expressions of HMGB1 and RAGE decreased,with statistical significance(P＜0.05).Compared with the high glucose+Gly group,the viability of H9c2 cells in the high glucose+Gly+pcDNA3.1-HMGB1 group decreased,the apoptosis rate,the number of autophagosomes and the expression of Bax,LC3 and Beclin-1 protein increased,the levels of IL-1β,TNF-α and ROS and the expression of HMGB1 and RAGE mRNA increased.The differences were statistically significant(P＜0.05).Conclusion:Gly can reduce the inflammation and oxidative stress response of H9c2 cells induced by high glucose,inhibit their apoptosis and autophagy,which may be related to the inhibition of HMGB1/RAGE signaling pathway.
glycyrrhizinhigh mobility group box proteinreceptor for advanced glycation end productshigh glucosecardiomyocytesapoptosisautophagyexperimental study
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维普
