FER-1 inhibits methylglyoxal-induced ferroptosis in mouse alveolar macrophages in vitro

doi:10.12122/j.issn.1673-4254.2024.12.21

Abstract

Abstract:

Objective To investigate the inhibitory effect of FER-1 on methylglyoxal-induced ferroptosis in cultured mouse alveolar macrophages. Methods MH-S cells derived from mouse alveolar macrophages treated with 90 μg/mL methylglyoxal, 10 μmol/mL FER-1MG+FER-1, or both were examined for intracellular reactive oxygen species (ROS), malondialdehyde (MDA) and ferrous ion (Fe²⁺) levels and changes in mitochondrial membrane potential. Western blotting was performed to detect the protein expression levels of glutathione peroxidase 4 (GPX4) and long-chain acyl-CoA synthase 4 (ACSL4). Results Methylglyoxal treatment of MH-S cells for 24 h significantly decreased the protein expression level of GPX4, upregulated the protein expression of ACSL4, increased intracellular concentrations of ferrous ions, ROS and MDA, caused loss of mitochondrial membrane potential, and decreased cell viability. Treatment of the cells with FER-1 effectively attenuated these detrimental effects of methylglyoxal in MH-S cells by increasing GPX4 expression, reducing ACSL4 expression and intracellular ferrous ions, ROS and MDA levels, and restoring the mitochondrial membrane potential. Conclusion Methylglyoxal can induce ferroptosis in MH-S cells in a dose-dependent manner, and FER-1 can rescue the cells from methylglyoxal-induced ferroptosis.

Key words: ferroptosis inhibitor ferroptin-1, mouse alveolar macrophages, methylglyoxal, ferroptosis

Qi ZHANG, Zezhao JI, Abai JIASHAER∙, Youda WANG, ABUDUXUKUER∙Abulimiti. FER-1 inhibits methylglyoxal-induced ferroptosis in mouse alveolar macrophages in vitro[J]. Journal of Southern Medical University, 2024, 44(12): 2443-2448.

Figures/Tables 6

Fig.1 Effects of different concentrations (0-120 μg/mL) of methylglyoxal (MG) for 24 h on protein expressions of GPX4 and ACSL4 in mouse alveolar macrophages (Mean±SD, n=5). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs control group.

Fig.2 FER-1 inhibits MG-induced changes in expressions of GPX4 and ACSL4 in mouse alveolar macrophages (Mean±SD, n=5). ##P<0.01 vs control group; *P<0.05, **P<0.01 vs 90 μg/mL MG group.

Fig.3 FER-1 ameliorates MG-induced increase of intracellular ROS production in MH-S cells (Mean±SD, n=4). ####P<0.0001 vs control group; ****P<0.0001 vs 90 μg/mL MG group.

Fig.4 FER-1 ameliorates MG-induced increase in lipid peroxidation levels in MH-S cells (Mean±SD, n=4). ###P<0.001 vs control group, ****P<0.0001 vs 90 μg/mL MG group.

Fig.5 FER-1 inhibits MG-induced loss of mitochondrial membrane potential (MMP) in MH-S cells. A: Fluorescent intensity of MMP (red) of MH-S cells treated with MG, FER-1 or both (Original magnification: ×40). B: Quantitative analysis of fluorescence intensity of MMP (Mean±SD, n=4). ####P<0.0001 vs control group, ***P<0.001 vs 90 μg/mL MG group.

Fig.6 FER-1 inhibits MG-induced increase of ferrous iron ions in MH-S cells (Mean±SD, n=4). ###P<0.001 vs control group, ***P<0.001 vs 90 μg/mL MG group.

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