Hypoxia/reoxygenation-induced upregulation of miRNA-542-5p aggravated cardiomyocyte injury by repressing autophagy
| Autor: | Xin Min, Ting-Ting Jiang, Da-Li You, Li Wang, Fei Wang, Shan-You Hu, Xiao Wu |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Cancer Research Myocardial Reperfusion Injury Autophagy-Related Protein 7 Flow cytometry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Western blot Downregulation and upregulation Lactate dehydrogenase medicine Autophagy Humans Myocytes Cardiac Viability assay Hypoxia Cells Cultured medicine.diagnostic_test Cell Biology Cell biology Up-Regulation MicroRNAs 030104 developmental biology chemistry Apoptosis 030220 oncology & carcinogenesis Stem cell |
| Zdroj: | Human cell. 34(2) |
| ISSN: | 1749-0774 |
| Popis: | MicroRNAs (miRNAs) and autophagy exert an important role in hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury. The current study aimed to explore the role of miRNA and autophagy in H/R-induced cardiomyocyte injury. Cardiomyocyte H9c2 was exposed to H/R to simulate H/R injury in vitro. The differentially expressed miRNAs were identified using quantitative RT-PCR (qPCR). Lactate dehydrogenase (LDH) activity was assayed to assess H/R injury. The role of miRNA and autophagy in regulating the viability and cell apoptosis was evaluated using cell counting kit-8 (CCK-8) assay, flow cytometry (FCM), and western blot. The autophagy activation was assessed through testing the number of light chain 3 (LC3) puncta and LC3-II expression using western blot and immunofluorescence analysis. In the present study, we found that the miR-542-5p expression and the autophagy activation were significantly increased in H9c2 cells after H/R injury. Functionally, forced expression of miR-542-5p further aggravated H/R injury in H9c2 cells, whereas miR-542-5p inhibition alleviated H/R injury as measured by the cell viability, LDH activity and cell apoptosis. miR-542-5p repressed autophagy activation, whereas miR-542-5p inhibition facilitated autophagy activation in H9c2 cells exposed to H/R as measured by the LC3 puncta number, LC3II, and p62 protein level. Especially, autophagy inhibition by specific inhibitor partially lessened the role of miR-542-5p inhibitor in alleviating H/R injury. Finally, the autophagy-related 7 (ATG7) was identified as a novel target gene of miR-542-5p in H9c2 cells. The current data suggest that miR-542-5p/autophagy pathway might be a potential target for the treatment of H/R-related heart diseases. |
| Databáze: | OpenAIRE |
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