FTOrepressesNLRP3‐mediated pyroptosis and alleviates myocardial ischemia–reperfusion injury via inhibitingCBL‐mediated ubiquitination and degradation of β‐catenin

Abstract

Cardiac ischemia/reperfusion (I/R) injury is a complicated pathological event, which has close association with pyroptosis. This study uncovered the regulatory mechanisms of fat mass and obesity-associated protein (FTO) in NLRP3-mediated pyroptosis during cardiac I/R injury. H9c2 cells were stimulated with oxygenglucose deprivation/reoxygenation (OGD/R). Cell viability and pyroptosis were detected by CCK-8 and flow cytometry. Western blotting or RT-qPCR was performed to analyze target molecule expression. NLRP3 and Caspase-1 expression was observed by immunofluorescence staining. IL-18 and IL-1β production was detected by ELISA. The total m6A and m6A level of CBL was determined by dot blot assay and methylated RNA immunoprecipitation-qPCR, respectively. The interaction between IGF2BP3 and CBL mRNA was confirmed by RNA pull-down and RIP assays. The protein interaction between CBL and βcatenin and βcatenin ubiquitination were evaluated by Co-IP. Myocardial I/R model was established in rats. We determined infarct size by TTC staining and pathological changes by H&E staining. LDH, CK-MB, LVFS, and LVEF were also assessed. FTO and βcatenin were down-regulated, while CBL was up-regulated by OGD/R stimulation. FTO/β-catenin overexpression or CBL silencing restrained OGD/R-induced NLRP3 inflammasome-mediated pyroptosis. CBL repressed βcatenin expression via ubiquitination and degradation. FTO reduced the mRNA stability of CBL by inhibiting m6A modification. CBL-mediated ubiquitination and degradation of βcatenin were involved in FTO-induced pyroptosis inhibition during myocardial I/R injury. FTO inhibits NLRP3-mediated pyroptosis to attenuate myocardial I/R injury via repressing CBL-induced ubiquitination degradation of βcatenin.

Fat mass and obesity associated protein inhibits neuronal ferroptosis via the FYN/Drp1 axis and alleviate cerebral ischemia/reperfusion injury

“…Earlier research discovered that FTO inhibits OS in cerebral I/R injury by mediating m6A modification of Nrf2 mRNA 30 . As a demethylase, FTO is noted to suppress m6A modification of proteins to afflict their stability 43,44 . Our data proved that FYN was upregulated after tMCAO/R treatment and FTO overexpression reduced FYN expression by decreasing FYN mRNA m6A modification.…”

“… 30 As a demethylase, FTO is noted to suppress m6A modification of proteins to afflict their stability. 43 , 44 Our data proved that FYN was upregulated after tMCAO/R treatment and FTO overexpression reduced FYN expression by decreasing FYN mRNA m6A modification. Considering that FYN may regulate the process of cerebral I/R, 14 we also designed related experiments to ascertain the specific effects of FYN on cerebral I/R injury.…”

“…19,42 Our experiments unveiled reduced Fe 2+ and 4HNE contents and enhanced GPX4 levels to afflict their stability. 43,44 Our data proved that FYN was upregulated after tMCAO/R treatment and FTO overexpression reduced FYN expression by decreasing FYN mRNA m6A modification.…”

An effective disease diagnostic model related to pyroptosis in ischemic cardiomyopathy

“…The pyroptosis‐related genes play essential roles in the occurrence and development of cardiovascular disease and affect the process and outcomes. Inhibiting myocardial pyroptosis could improve cardiac remodelling 44 . This study confirmed 13 optimal pyroptosis genes to be downregulated or upregulated in ICM, demonstrating a necessary diagnostic and regulating potential.…”

“…Inhibiting myocardial pyroptosis could improve cardiac remodelling. 44 This study confirmed 13 optimal pyroptosis genes to be downregulated or upregulated in ICM, demonstrating a necessary diagnostic and regulating potential. We identified these genes as highly accurate biomarkers for the effective diagnosis of ICM enabling early treatment and intervention.…”