Metformin ameliorates ferroptosis in cardiac ischemia and reperfusion by reducing NOX4 expression via promoting AMPKα

• Published in: Pharmaceutical biology Vol. 61; no. 1; pp. 886 - 896
• Main Authors: Wu, Zhenhua, Bai, Yunpeng, Qi, Yujuan, Chang, Chao, Jiao, Yan, Bai, Yaobang, Guo, Zhigang
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 01-12-2023; Taylor & Francis Ltd; Taylor & Francis Group
• Subjects: Adenosine kinase; AMP; AMP-Activated Protein Kinases - metabolism; Animals; Apoptosis; Cell Line; Cell viability; Cholecystokinin; Enzyme-linked immunosorbent assay; Ferroptosis; Fluorescein diacetate; Gene expression; Glucose; Heme; Immunohistochemistry; Iron - metabolism; Ischemia; Kinases; LDH; MDA; Metformin; Metformin - pharmacology; Mitochondrial damage; Myocardial Ischemia - metabolism; Myocytes, Cardiac; non-heme iron; NOX4 protein; Oxygen; Polymerase chain reaction; Rats; Rats, Sprague-Dawley; Reperfusion; Reperfusion Injury - drug therapy; Reperfusion Injury - metabolism; Reperfusion Injury - prevention & control; Reverse transcription; ROS; siRNA; LDH; non-heme iron; MDA; Mitochondrial damage; ROS
• ISSN: 1388-0209; 1744-5116
• DOI: 10.1080/13880209.2023.2212700

Metformin (Met) has a protective effect against cardiac ischemia and reperfusion (I/R) injury.
This study uncovered the Met effect on ferroptosis in cardiac I/R.
Sprague-Dawley rats underwent cardiac I/R treatment (ischaemia 30 min; reperfusion 24 h) (I/R group), and administered intravenously with Met (200 mg/kg) (I/R + Met group). Haematoxylin-eosin staining, Prussian blue staining, immunohistochemistry and transmission electron microscope were conducted on cardiac tissues. H9c2 cells underwent oxygen-glucose deprivation/reoxygenation (OGD/R group) and treated by Met (0.1 mM) (OGD/R + Met group). Adenosine monophosphate-activated protein kinase α (AMPKα) siRNA was transfected into OGD/R-induced H9c2 cells. Cell counting kit-8 (CCK-8) assay, dichloro-dihydro-fluorescein diacetate (DCFH-DA) and JC-1 staining were conducted on H9c2 cells. Ferroptosis-related indicators and gene expression were detected by enzyme-linked immunosorbent assay (ELISA), quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot.
In cardiac I/R rat, Met decreased heart and serum MDA, cardiac and serum non-heme iron, and serum CK-MB and LDH (inhibition rate: 50.0%, 48.8%, 47.6%, 29.5%, 30.6% and 34.7%, respectively), relieved cardiac tissue ferroptosis and mitochondria damage, increased fraction shortening and ejection fraction (157.5% and 146.2% on day 28, respectively), up-regulated AMPKα and down-regulated NOX4 in cardiac tissues. In OGD/R-induced H9c2 cells, Met (0.1 mM) increased cell viability (promotion rate: 170.0%), decreased non-heme iron and MDA (inhibition rate: 30.1% and 47.9%, respectively), relieved ferroptosis, up-regulated AMPKα and down-regulated NOX4. AMPKα silencing abrogated these effects of Met on the OGD/R-induced H9c2 cells.
Met shows effectiveness in relieving ferroptosis in cardiac I/R. In the future, Met may be an effective drug for relieving ferroptosis in cardiac I/R patients clinically.