Mechanism of miR-370-3p/SESN2 regulating lipopolysaccharide-induced apoptosis of mouse alveolar epithelial cells

Wu Zhouyou, Li Ting, Zhang Tengwei, Fang Qiaoyan, Yang Liu, Li Qiao

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Journal of Chongqing Medical University ›› 2024, Vol. 49 ›› Issue (09) : 1121-1128. DOI: 10.13406/j.cnki.cyxb.003590
Basic research

Mechanism of miR-370-3p/SESN2 regulating lipopolysaccharide-induced apoptosis of mouse alveolar epithelial cells

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Abstract

Objective To investigate the mechanism of miR-370-3p/Sestrin2（SESN2） regulating the apoptosis of mouse alveolar epithelial cells（AECs） induced by lipopolysaccharide（LPS）. Methods C57BL/6J mice were randomly divided into LPS+anti-NC group and LPS+miR-370-3p inhibitor（anti-miR-370-3p） group，with 12 mice in each group. LPS（5 mg/kg） was given by intratracheal injection to induce acute lung injury（ALI），and anti-miR-370-3p or anti-NC was injected via the caudal vein of mice. MLE12 cells were randomly divided into control group（Con+anti-NC），Con+anti-miR-370-3p group，LPS+anti-NC group，LPS+anti-miR-370-3p group，and LPS+si-SESN2+anti-miR-370-3p group. Quantitative real-time PCR was used to analyze miR-370-3p in lung tissue or cells. Mitochondrial morphology and mitochondrial membrane potential were measured to investigate the protective effect of miR-370-3p knockdown against LPS-induced mitochondrial dysfunction. Dual-luciferase reporter assay confirmed the targeting relationship between miR-370-3p and SESN2. Results Compared with the LPS+anti-NC group，the LPS+anti-miR-370-3p group had significant reductions in the expression of miR-370-3p and the protein expression levels of receptor-interacting serine/threonine-protein kinase 3（RIPK3） and mixed lineage kinase domain-like（MLKL） in lung tissue，as well as a significant reduction in lung inflammation score（P<0.05） and a significant increase in the protein expression level of surfactant protein C（SPC） in lung tissue（P<0.001）. Compared with the LPS+anti-NC group，the LPS+anti-miR-370-3p group had significant reductions in the protein expression levels of RIPK3 and MLKL in MLE12 cells and the percentage of PI-positive cells（P<0.05），as well as significant increases in mitochondrial fragmentation and mitochondrial membrane potential（P<0.05）. MiR-370-3p could inhibit the luciferase activity of SESN2-WT reporter gene. Compared with the LPS+anti-miR-370-3p group，the LPS+Si-SEN2+anti-miR-370-3p group had significant reductions in mitochondrial fragmentation and mitochondrial membrane potential（P<0.05）. Conclusion The miR-370-3p/SESN2 axis promotes the necrotizing apoptosis of AECs during ALI induced by LPS by mediating mitochondrial breakage and damaging mitochondrial function.

Key words

miR-370-3p / Sestrin2 / lipopolysaccharide / alveolar epithelial cells / necrotizing apoptosis

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Wu Zhouyou , Li Ting , Zhang Tengwei , et al . Mechanism of miR-370-3p/SESN2 regulating lipopolysaccharide-induced apoptosis of mouse alveolar epithelial cells. Journal of Chongqing Medical University. 2024, 49(09): 1121-1128 https://doi.org/10.13406/j.cnki.cyxb.003590

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