Glycyrrhetinic Acid Ameliorates Sorafenib-induced Cardiomyocyte Injury and Apoptosis through HIF-1α/BNIP3-mediated Mitophagy

doi:10.3870/j.issn.1672-0741.24.12.008

Abstract

Abstract: Objective The effects of glycyrrhetinic acid on sorafenib-induced cardiomyocyte injury and apoptosis were investigated via the hypoxia-inducible factor-1α(HIF-1α)/adenovirus E1B 19 kDa-interacting protein 3(BNIP3)signaling pathway. Methods MTT was used to determine the effects of glycyrrhetinic acid on H9c2 cells in rat cardiomyocytes.The experiment was divided into control group,sorafenib group,sorafenib+glycyrrhetinic acid group,sorafenib+glycyrrhetinic acid+HIF-1α inhibitor YC-1 group,and sorafenib+glycyrrhetinic acid+YC-1+autophagy agonist rapamycin(RAPA)group.A kit was used to determine LDH release and MDA levels.The fluorescent probe method was used to determine the level of total ROS in the cells.Apoptosis was detected via flow cytometry.Immunofluorescence was used to detect the protein expression of LC3 and the colocalization of BNIP3 and LC3.The expression levels of the mitochondrial autophagy-related proteins LC3 II/I,p62 and Beclin-1 were detected via Western blotting.RT-qPCR and Western blotting were used to detect the mRNA and protein expression levels of HIF-1α and BNIP3,respectively. Results Compared with that in the sorafenib group,the viability of H9c2 cells in the 100 and 200 μmol/L glycyrrhetinic acid groups was significantly increased(P<0.05).Compared with those in the control group,LDH release and the MDA and ROS levels in H9c2 cells in the sorafenib group were significantly up-regulated(P<0.05);the apoptosis rate of cells was significantly increased(P<0.05);the expression levels of LC3 II/I and Beclin-1 protein were significantly down-regulated(P<0.05);and the levels of p62 were significantly up-regulated(P<0.05);the fluorescence intensity of LC3 in H9c2 cells was weakened,and the co-localization of LC3 and BNIP3 in H9c2 cells was reduced,the mRNA and protein expression levels of HIF-1α and BNIP3 were significantly down-regulated(P<0.05).Compared with those in the sorafenib group,LDH release and the MDA and ROS levels in H9c2 cells in the sorafenib+glycyrrhetinic acid group were significantly down-regulated(P<0.05);the apoptosis rate of H9c2 cells was significantly reduced(P<0.05);the expression levels of LC3 II/I and Beclin-1 proteins were significantly up-regulated(P<0.05);the p62 levels were significantly down-regulated(P<0.05);the fluorescence intensity of LC3 in H9c2 cells was enhanced,the colocalization of LC3 and BNIP3 in H9c2 cells was enhanced,the mRNA and protein expression levels of HIF-1α and BNIP3 were significantly up-regulated(P<0.05).YC-1 significantly reversed these effects(P<0.05),and RAPA significantly inhibited the effect of YC-1(P<0.05). Conclusion Glycyrrhetinic acid can promote mitophagy by activating the HIF-1α/BNIP3 signaling pathway and can inhibit sorafenib-induced cardiomyocyte injury and apoptosis.

Key words: glycyrrhetinic acid, HIF-1α/BNIP3, mitophagy, sorafenib, cardiomyocyte

CLC Number:

R542.2

Zhang Xianlin, Wang Lizhen, Zhang Jie et al. Glycyrrhetinic Acid Ameliorates Sorafenib-induced Cardiomyocyte Injury and Apoptosis through HIF-1α/BNIP3-mediated Mitophagy[J]. Acta Medicinae Universitatis Scientiae et Technologiae Huazhong, 2026, 55(1): 68-75.

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