MiR-448-5p/VEGFA Axis Protects Cardiomyocytes from Hypoxia Through Regulating the FAS/FAS-L Signaling Pathway

Abstract

Bioinformatics analysis showed that miR-448-5p expression in the myocardial tissue of rats with myocardial infarction significantly increased, suggesting that it may participate in myocardial cell apoptosis in myocardial infarction. This study aimed to explore the protective effects of miR-448-5p on hypoxic myocardial cells.H9C2 cells were cultured and subjected to anoxia for 2, 4, and 8 hours to establish a hypoxia model. MiR-448-5p mimic and inhibitor were transfected into the cells; then, a dual-luciferase experiment was conducted to verify the targeting relationship between miR-448-5p and VEGFA. Cell viability and apoptosis was detected by cell counting kit-8 and flow cytometry, respectively. The expressions of apoptosis-related proteins, miR-448-5p, FAS, and FAS-L were measured using western blotting and quantitative reverse transcription-polymerase chain reaction (qRT-PCR).Hypoxia-reduced H9C2 cell viability and promoted apoptosis. MiR-448-5p expression was increased after H9C2 cell hypoxia. MiR-448-5p mimic significantly inhibited the viability and promoted the apoptosis of hypoxia-induced model cells. Hypoxia promoted the expression of apoptosis-related protein B-cell lymphoma-2 (Bcl-2) and inhibited the expressions of Bcl-2-associated x protein (Bax), cleaved caspase-3, and caspase-3, whereas the effect of inhibitor on hypoxia-reduced H9C2 cell and apoptotic protein expression were opposite to miR-448-5p mimic. MiR-448-5p targeted VEGFA and regulated its expression. Silenced VEGFA expression significantly inhibited inhibitor effect on increasing cell viability and promoted apoptosis. In addition, miR-448-5p mimic inhibited the effect of hypoxia on promoting the expressions of FAS and FAS-L of H9C2 cells. Inhibitors had the opposite effect on cell hypoxia model.The miR-448-5p/VEGFA axis could protect cardiomyocytes from hypoxia through inhibiting the FAS/FAS-L signaling pathway.

Analysis of the role and mechanism of EGCG in septic cardiomyopathy based on network pharmacology

“…However, miR-15a-5p suppresses the inflammatory response of human peritoneal interstitial cells by targeting VEGFA ( Shang et al, 2018 ). VEGFA promotes cardiomyocyte apoptosis induced by hypoxia, and miR-448-5p targeting VEGFA can inhibit the expression of Caspase3 ( Tang et al, 2021 ). The binding energy of EGCG and VEGFA is 5.45 kcal/mol.…”

Induction of ligustrazine apoptosis of A549 cells through activation of death receptor pathway

“…Both vivo and in vitro studies have demonstrated that Fas mAb can bind to Fas, resulting in the death of cells expressing Fas. Thus Fas, as a target, is considered a key factor in cell death (19). High expression of Fas on the cell membrane is thus highly indicative of the susceptibility of Fas signaling pathway to induce apoptosis; however, the cutoff value between tumor cells is not uniform.…”

Transient Focal Cerebral Ischemia Leads to miRNA Alterations in Different Brain Regions, Blood Serum, Liver, and Spleen

“…For miR-448, no alteration in the brain following stroke was reported, nor a connection to ischemic stroke and blood levels. A study utilizing a spinal cord ischemia/reperfusion injury model reported increased levels of miR-448 measured by qRT-PCR [ 44 ] and in cell hypoxic culture models of myocardial infarction [ 31 , 45 ]. The studies, as mentioned above, on miRNA alteration in ischemic stroke have either been performed in the cerebral cortex, the blood serum, or the researchers did not indicate the exact location.…”

“…The last miRNA, miR-448-5p, was also scarcely studied until now. One study found VEGFA (vascular endothelial growth factor A) to be a target and a regulator of the FAS/FAS-L signaling pathway in a hypoxia model in myocardial rat cells [ 31 ]. VEGFA is known for angiogenesis and neuroprotection but seems to have a dual role since it alters vascular permeability and blood–brain barrier integrity in stroke [ 32 , 33 ].…”