Exosomes Derived from Anoxia Preconditioned Mesenchymal Stem Cells alleviate Myocardial Ischemia Reperfusion Injury by Inhibiting Pyroptosis/ Caspase-1 Induced Apoptosis in The Deficiency of Gasdermin D
Abstract BackgroundExosomes derived from adipose-derived mesenchymal stem cells can potentially protect cardiomyocytes from myocardial ischemia reperfusion injury. It's notable that exosomes derived from adipose-derived mesenchymal stem cells underwent anoxia preconditioning showed a better cardioprotective effect than that without anoxia. Here, in vitro and in vivo studies were used to investigate the cardioprotective effects against myocardial ischemia reperfusion injury of exosomes derived from adipose-derived mesenchymal stem cells with (Int-EXO) or without anoxia (NC-EXO), respectively. Methods: Adipose-derived mesenchymal stem cells and exosomes were identified by western blot, flow cytometry, transmission electron microscopy, and nanosight. An exosome tracer assay identified exosomes absorbed by cells. An in vitro model using mice cardiomyocytes for studying anoxia-reoxygenation and an in vivo mice model of MIRI were used to investigate the cardioprotective effects of NC-Exo and Int-Exo, respectively.ResultsWe discovered that treatment with NC-EXO and Int-EXO significantly reduced the infarct size and attenuated cardiomyocyte apoptosis, In addition, Int-EXO group had a less infarct size and apoptosis degree. The mechanism revealed by RNA sequencing showed that 40 miRNAs were up-regulated in Int-EXO compared to NC-EXO. 10 of these miRNAs could bind thioredoxin-interacting protein as a downstream target gene; among these, the top-discrepant miRNA224-5p was selected for further study. Dual luciferase reporter assay and rescue study verified TXNIP as a target gene for miR-224-5p. Furthermore, the cellular death signaling pathway which Int-EXO involved in mediating was in a special form of apoptosis, not pyroptosis, induced by activated thioredoxin-interacting protein-pyroptosis-caspase1 pathway in gasdermin D-deficient cells. ConclusionThe research demonstrated adipose-derived mesenchymal stem cells exosomes attenuated MIRI by inhibiting pyroptosis-induced apoptosis in cardiomyocytes which are lack of gasdermin D. The cardioprotective effect of Int-EXO was more significant than that of NC-EXO, possibly due to treated with anoxia preconditioning, adipose-derived mesenchymal stem cells product more miRNAs targeting thioredoxin-interacting protein in exosomes to alleviate pyroptosis-induced apoptosis. These findings provide new insights into the pathogenesis and methods for intervention of myocardial ischemia reperfusion injury.
