| Objective: Anthracycline is one of the most effective drugs for the treatment of diseases such as leukemia and lymphoma,but its clinical application is limited by cardiotoxicity due to its dose accumulation.With the breakthrough in the treatment of hematological malignancy,the clinical use of daunorubicin(DNR)is increasingly demanded.However,current myocardial protective drugs,such as dexrazoxane(DZR),fail to completely antagonize anthracycline cardiotoxicity.Mesenchymal stem cells(MSCs)are a class of stem cells capable of self-renewal and multi-lineage differentiation into cells of mesoderm and embase immunomodulatory ability.Our study was to investigate effects of mesenchymal stem cells on the daunorubicin-induced myocardial injury and how to enhance the cardioprotective effect of mesenchymal stem cells.Methods:1 The hBMSCs were centrifuged and cultured in a plastic culture flask,then morphology,immunophenotype and multi-lineage differentiation of passage-3 cells were identified.2 Build rat models of subacute or chronic daunorubicin-derived myocardial injury by DNR 4mg/kg i.v per 48 h for six times lasting for 28 days or DNR 2mg/kg qw for six times by 8 weeks.3 To explore the protection of BMSCs on subacute myocardial injury induced by DNR,54 SD rats were randomly divided into 9 groups,6 rats in each group.BMSCs and the following drugs were injected through the tail vein,DNR with 4mg/kg i.v q48 h for 6 times,DZR 80mg/kg i.v q48 h for 6 times and 30 mins parallelly before DNR,and 1×106 BMSCs once.Groups are divided as follows: Control groups:(1)blank group(blank Control group): given equal volume of 0.9% saline.(2)Dexrazoxane group(DZR group): DZR only.(3)Mesenchymal stem cell group(MSC group): BMSC only.Experimental groups:(4)Daunorubicin group(DNR group): DNR only.(5)DZR + DNR group(DD group): receiving DZR 30 mins before DNR.(6)Mesenchymal stem cells 3 days pre-infusion+DNR group(-3MD group): MSCs infusion 3 days before the first dose of DNR.(7)Mesenchymal stem cells 3 days pre-infusion+DZR+DNR group(-3MDD group): MSCs infusion 3 days before the first dose of DNR,along with parallel DZR.(8)Mesenchymal stem cells 7 days post-infusion+DNR group(7MD group): MSCs infusion 7 days after the first dose of DNR;(9)Mesenchymal stem cells 7 days post-infusion+DZR+DNR group(7MDD group): MSCs infusion 7 days after the first dose of DNR,along with parallel DZR.Rats were anesthetized by 50mg/kg pentobarbital and sacrificed after ultrasonic echocardiography examination on the 28 th day.Heart tissues was examined by H&E stain and the expression of CD3 and HLA-DR surface antigens by immunohistochemistry stain.Results:1 Passage-3 mesenchymal stem cells showed highly consistent,long spindle-shaped and significant round or oval nuclei inside with loose cytoplasm and rich particles.The amounts of negative expressed CD31,CD34,CD45 and positive expressed CD29,CD44,CD73 on mesenchymal stem cells were 0.3%,0.6%,0.2%,96.8%,99.9% and 96.7%.Then we chose highly expressed CD44 and CD73 and negative CD34 and CD45 as identification markers.Also MSCs were well induced towards osteoblasts,adipocytes and chondrocytes in corresponding inducible culture flasks.2 We succeeded in building subacute DNR-induced myocardial injury model by DNR 4mg/kg i.v q48 h for 6 times as heart tissue damage and impaired heart function were examed at the 28 th day.While chronic DNR-induced myocardial injury model by DNR 4mg/kg i.v q48 h for 6 times came out with less cardiotoxicity and more drug-induced hepatotoxicity.So we chose subacute DNR-induced myocardial injury model for further study.3 Compared with Blank group,DZR and MSC groups made no differences,while DNR induced a obvious myocardial injury marked by dramatical decrease in values of ejection fraction(EF)and fraction shortening(FS)in DNR group(P<0.05).Compared with DNR group,DZR succeeded in increasing values of EF and FS,resulting in improving the heart function(P<0.05).For two different time points of MSCs treatment to DNR induced myocardiotoxicity,we observered that MSCs 3 days pre-infusion or combined with DZR significantly increased values of EF and FS,compared with DNR group(P<0.05).MSCs 7 days post-infusion combined with DZR could also significantly increase values of EF and FS(P<0.05).However,there was no significant difference in the left ventricular systolic and diastolic function in 7MD group(P>0.05).We also observed that there was no T cell and antigen-presenting cell(APC)infiltration in the Blank,DZR and MSC groups.Infiltrations of T cells and APC cells were observed in DNR group.Compared with DNR group,infiltrations of T cells and APC cells in DD group,-3MD group,-3MDD group and 7MDD group were significantly decreased,but not dramatically decrease in-7MD group(P>0.05).Compared with DD group,3MD and-3MDD groups could further reduce the infiltration of T cells and APC cells.Conclusion:1 We succeeded in isolating human bone marrow-derived mesenchymal stem cells,through identification of morphology,immunophenotype and multi-lineage differentiation.2 We succeeded in building subacute daunorubicin-induced myocardial injury model in rats by a protocol of DNR 4mg/kg i.v q48 h for 6 times.3 A three-day pre-infusion of mesenchymal stem cells can be more effective to prevent myocardial injury from daunorubicin.MSCs combined with DZR can further enhance the therapeutic effect,but there are individual differences.4 MSCs may reduce myocardial damage and improve cardiac function by suppressing the infiltration of T cells and antigen-presenting cells. |
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