Effect And Mechanism Of Tongxinluo Drug Pretreatment Of Bone Marrow Mesenchymal Stem Cells On Myocardial Repair After Acute Myocardial Infarctio

Tongxinluo-pretreated mesenchymal stem cells facilitate cardiac repair via exosomal transfer of miR-146a-5p targeting IRAK1/NF-κB p65 pathwayBackground:Bone marrow cells(BMCs),especially mesenchymal stem cells(MSCs)have shown attractive application prospects in acute myocardial infarction(AMI).However,the weak efficacy becomes their main limitation in clinical translation.Based on the anti-inflammation and anti-apoptosis effects of a Chinese medicine-Tongxinluo(TXL),we aimed to explore the effects of TXL-pretreated MSCs(MSCsTXL)in enhancing cardiac repair and further investigated the underlying mechanism.Methods:MSCsTXL or MSCs and the derived exosomes(MSCsTXL-exo or MSCs-exo)were collected and injected into the infarct zone of rat hearts.In vivo,the anti-apoptotic and anti-inflammation effects,and cardiac functional and histological recovery were evaluated.In vitro,the apoptosis was evaluated by western blotting and flow cytometry.miRNA sequencing was utilized to identify the significant differentially expressed miRNAs between MSCsTXL-exo and MSCs-exo,and the miRNA mimics and inhibitors were applied to explore the specific mechanism.Results:Compared to MSCs,MSCsTXL enhanced cardiac repair with reduced cardiomyocytes apoptosis and inflammation at the early stage of AMI,and significantly improved left ventricular ejection fraction(LVEF)with reduced infarct size in an exosome-dependent way.Similarly,MSCsTXL-exo exerted superior therapeutic effects in anti-apoptosis and anti-inflammation,as well as improving LVEF and reducing infarct size compared to MSCs-exo.Further exosomal miRNA analysis demonstrated that miR-146a5p was the candidate effector of the superior effects of MSCsTXL-exo.Besides,miR-146a5p targeted and decreased IRAK1,which inhibited the nuclear translocation of NF-κB p65 thus protecting H9C2 cells from hypoxia injury.Conclusions:This study suggested that MSCsTXL markedly facilitated cardiac repair via a new mechanism of the exosomal transfer of miR-146a-5p targeting IRAK1/NF-κB p65 pathway,which has great potential for clinical translation.Sequential transplantation of exosomes and mesenchymal stem cells pretreated with a combination of hypoxia and Tongxinluo efficiently facilitates cardiac repairBackground:Bone marrow-derived mesenchymal stem cells(MSCs),which possess immunomodulatory characteristic,are promising candidates for the treatment of acute myocardial infarction(AMI).However,the low retention and survival rate of MSCs in the ischemic heart limit their therapeutic efficacy.Strategies either modifying MSCs or alleviating the inflammatory environment,which facilitates the recruitment and survival of the engrafted MSCs,may solve the problem.Thus,we aimed to explore the therapeutic efficacy of sequential transplantation of exosomes and combinatorial pretreated MSCs in the treatment of AMI.Methods:Exosomes derived from MSCs were delivered to infarcted hearts through intramyocardial injection followed by the intravenous infusion of differentially pretreated MSCs on Day 3 post-AMI.Enzyme linked immunosorbent assay(ELISA)was performed to evaluate the inflammation level as well as the SDF-1 levels in the infarcted border zone of the heart.Echocardiography and histological analysis were performed to assess cardiac function,infarct size,collagen area and angiogenesis.Results:Sequential transplantation of exosomes and the combinatorial pretreated MSCs significantly facilitated cardiac repair compared to AMI rats treated with exosomes alone.Notably,compared to the other three methods of co-transplantation,combinatorial pretreatment with hypoxia and Tongxinluo(TXL)markedly enhanced the CXCR4 level of MSCs and promoted recruitment,which resulted in better cardiac function,smaller infarct size and enhanced angiogenesis.We further demonstrated that exosomes effectively reduced apoptosis in MSCs in vitro.Conclusion:Sequential delivery of exosomes and pretreated MSCs facilitated cardiac repair post-AMI,and combined pretreatment with hypoxia and TXL better enhanced the cardioprotective effects.This method provides new insight into the clinical translation of stem cell-based therapy for AMI.