Department of Cardiology, Affiliated Haikou Hospital of Xiangya Medical College, Central South University, Haikou, China.
Department of Cardiology, Affiliated Haikou Hospital of Xiangya Medical College, Central South University, Haikou, China -
Panminerva Med. 2024 Sep;66(3):249-255. doi: 10.23736/S0031-0808.20.03979-8. Epub 2020 Jul 27.
The aim of this study was to investigate the effect of microRNA-30d (miR-30d) on autophagy and reveal the mechanism of autophagy promoting ferroptosis in H9C2 cells.
First, we detected miR-30d expression of myocardial tissue in the sham and myocardial infarction (MI) group, and then analyzed by biochemical analysis and luciferase Genetic experiments to confirm its downstream target gene of. After using Lentivirus-ATG5 (LV-sh-ATG5) to effectively inhibit autophagy, in order to further clarify the possible mechanism of autophagy leading to ferroptosis in H9C2 cells, we have tested the relevant indicators ferroptosis.
We first found that miR-30d expression was down-regulated in myocardial tissue after MI, while autophagy increased, and autophagy was reduced when miR-30d was overexpressed, and then analyzed by biochemical analysis and luciferase Genetic experiments confirmed that ATG5 was a downstream target gene of miR-30d. After using Lentivirus-ATG5 (LV-shATG5) to effectively inhibit autophagy and up-regulate the expression of FTH1 and GSH peroxidase (GPX4) in H9C2 cells, reduce the content of MDA, increase the content of GSH, and increase the activity of GPX4, suggesting that autophagy after MI may promote ferroptosis in H9C2 cells.
The expression of miR-30d decreased in cardiomyocytes after MI and which can inhibit autophagy by binding to ATG5. Furthermore, autophagy after MI may promote ferroptosis.
本研究旨在探讨 microRNA-30d(miR-30d)对自噬的影响,并揭示自噬促进 H9C2 细胞铁死亡的机制。
首先,我们检测了假手术和心肌梗死(MI)组心肌组织中 miR-30d 的表达,然后通过生化分析和荧光素酶遗传实验分析来确认其下游靶基因。在用 Lentivirus-ATG5(LV-sh-ATG5)有效抑制自噬后,为了进一步阐明自噬导致 H9C2 细胞铁死亡的可能机制,我们测试了铁死亡的相关指标。
我们首先发现 MI 后心肌组织中 miR-30d 的表达下调,而自噬增加,过表达 miR-30d 后自噬减少,然后通过生化分析和荧光素酶遗传实验分析证实 ATG5 是 miR-30d 的下游靶基因。用 Lentivirus-ATG5(LV-shATG5)有效抑制自噬并上调 H9C2 细胞中 FTH1 和谷胱甘肽过氧化物酶(GPX4)的表达,降低 MDA 含量,增加 GSH 含量,增加 GPX4 活性,表明 MI 后自噬可能促进 H9C2 细胞铁死亡。
MI 后心肌细胞中 miR-30d 的表达下调,可通过与 ATG5 结合抑制自噬。此外,MI 后自噬可能促进铁死亡。