Department of Pathology, University of Cincinnati, 231-Albert Sabin Way, Cincinnati, OH 45267, USA.
J Mol Med (Berl). 2012 Sep;90(9):997-1010. doi: 10.1007/s00109-012-0920-1. Epub 2012 May 31.
This study seeks to test our hypothesis that transgenic induction of miR-210 in mesenchymal stem cells (MSC) simulates the pro-survival effects of ischemic preconditioning (IPC) and that engraftment of (PC)MSC helps in the functional recovery of ischemic heart by miR-210 transfer to host cardiomyocytes through gap junctions. miR-210 expression in MSC was achieved by IPC or nanoparticle-based transfection of miR-210 plasmid ((miR)MSC) and functional recovery of the infarcted heart of rat transplanted with (PC)MSC or (miR)MSC was evaluated. Both (PC)MSC and (miR)MSC showed higher survival under lethal anoxia as compared to (non-PC)MSC and scramble-transfected MSC ((Sc)MSC) controls with concomitantly lower CASP8AP2 expression. Similarly, both (PC)MSC and (miR)MSC survived better and accelerated functional recovery of ischemic heart post-transplantation. To validate our hypothesis that MSC deliver miR-210 to host cardiomyocytes, in vitro co-culture between cardiomyocytes and (PC)MSC or (miR)MSC (using (non-PC)MSC or (Sc)MSC as controls) showed co-localization of miR-210 with gap-junctional connexin-43. miR-210 transfer to cardiomyocytes was blocked by heptanol pretreatment. Moreover, higher survival of cardiomyocytes co-cultured with (PC)MSC was observed with concomitant expression of CASP8AP2 as compared to cardiomyocytes co-cultured with (non-PC)MSC thus suggesting that miR-210 was translocated from MSC to protect host cardiomyocytes. Induction of miR-210 in MSC promoted their survival post-engraftment in the infarcted heart. Moreover, direct transfer of pro-survival miR-210 from (miR)MSC to host cardiomyocytes led to functional recovery of the ischemic heart.
本研究旨在验证我们的假设，即转染 miR-210 可模拟间充质干细胞（MSC）的缺血预适应（IPC）的生存效应，并且 MSC 移植有助于缺血性心脏的功能恢复，因为 miR-210 通过缝隙连接传递给宿主心肌细胞。通过 IPC 或基于纳米粒子的 miR-210 质粒转染（miR-MSC）实现 MSC 中的 miR-210 表达，并评估移植（PC）MSC 或（miR）MSC 的大鼠梗死心脏的功能恢复。与（非-PC）MSC 和 scramble 转染的 MSC（Sc-MSC）对照相比，（PC）MSC 和（miR）MSC 在致命缺氧下表现出更高的存活率，同时 CASP8AP2 表达降低。同样，（PC）MSC 和（miR）MSC 移植后均可更好地存活并加速缺血性心脏的功能恢复。为了验证我们的假设，即 MSC 将 miR-210 递送至宿主心肌细胞，体外共培养心肌细胞与（PC）MSC 或（miR）MSC（以（非-PC）MSC 或（Sc）MSC 作为对照）显示 miR-210 与缝隙连接连接蛋白-43 共定位。用庚醇预处理可阻断 miR-210 向心肌细胞的转移。此外，与与（非-PC）MSC 共培养的心肌细胞相比，与（PC）MSC 共培养的心肌细胞观察到更高的存活率，同时伴随着 CASP8AP2 的表达，这表明 miR-210 从 MSC 转移以保护宿主心肌细胞。在梗死心脏中 MSC 中的 miR-210 诱导促进了其移植后的存活。此外，直接从（miR）MSC 向宿主心肌细胞传递生存促进的 miR-210 可导致缺血性心脏的功能恢复。
