Mesenchymal stem cells improve cardiac conduction by upregulation of connexin 43 through paracrine signaling
Mesenchymal stem cells improve cardiac conduction by upregulation of connexin 43 through paracrine signaling
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DOI：
10.1152/ajpheart.00533.2012
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发表时间：
2013-02-01
影响因子：
4.8
通讯作者：
Banach, Kathrin
中科院分区：
文献类型：
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作者：
Mureli, Shwetha;Gans, Christopher P.;Banach, Kathrin
Mureli S，Gans CP，Bare DJ，Geenen DL，Kumar NM，Banach K.间充质干细胞通过旁分泌信号上调连接蛋白43改善心脏传导。Am J Physiol Heart Circ Physiol 304：H600-H609，2013。首次发表于2012年12月15日; doi：10.1152/ajpheart.00533.2012.-间充质干细胞（MSCs）被证明可以提高细胞存活率，并减轻心脏心律失常时，移植到心脏组织，但是，很少有人知道的机制，其中MSCs修改心脏组织的电生理特性。我们的目的是区分心肌细胞和间充质干细胞之间的细胞-细胞偶联的影响，从MSC衍生的旁分泌因子对多细胞心肌细胞制剂的自发活动和传导速度（θ）。将HL-1细胞接种在微电极阵列上，并从场电位记录确定其自发活动和θ。在异细胞培养的MSCs和HL-1细胞中，伴随着MSCs和心肌细胞之间的细胞间偶联，搏动频率减弱（t（0 h）：2.26 +/-0.18Hz; t（4 h）：1.98 +/-0.26Hz; P < 0.01）。在补充MSC条件培养基（ConM）或酪氨酸（ConT）的HL-1单层细胞中，θ以时间依赖性方式显著增加（ConT：t（0 h）：2.4 cm/s +/- 0.2; t（4 h）：3.1 +/- 0.4 cm/s），而搏动频率保持恒定。ConM或ConT处理后，连接蛋白（Cx）43 mRNA和蛋白表达水平也在同一时间段内增加。ConT处理后低密度脂蛋白受体相关蛋白6（LRP 6）磷酸化增强暗示了Wnt信号通路。抑制MSC的Wnt分泌（IWP-2; 5 μ mol/l）降低了ConT诱导磷酸化LRP 6和增加θ的功效。β-连环蛋白（小豆蔻素; 10 μ mol/l）或GSK 3-α/β（LiCl; 5 mmol/l）的抑制也抑制了θ的变化，进一步支持MSC介导的Cx43上调部分通过分泌的Wnt配体和经典Wnt信号通路的激活发生的假设。
Mureli S, Gans CP, Bare DJ, Geenen DL, Kumar NM, Banach K. Mesenchymal stem cells improve cardiac conduction by upregulation of connexin 43 through paracrine signaling. Am J Physiol Heart Circ Physiol 304: H600-H609, 2013. First published December 15, 2012; doi:10.1152/ajpheart.00533.2012.-Mesenchymal stem cells (MSCs) were shown to improve cell survival and alleviate cardiac arrhythmias when transplanted into cardiac tissue; however, little is known about the mechanism by which MSCs modify the electrophysiological properties of cardiac tissue. We aimed to distinguish the influence of cell-cell coupling between myocytes and MSCs from that of MSC-derived paracrine factors on the spontaneous activity and conduction velocity (theta) of multicellular cardiomyocyte preparations. HL-1 cells were plated on microelectrode arrays and their spontaneous activity and theta was determined from field potential recordings. In heterocellular cultures of MSCs and HL-1 cells the beating frequency was attenuated (t(0h): 2.26 +/- 0.18 Hz; t(4h): 1.98 +/- 0.26 Hz; P < 0.01) concomitant to the intercellular coupling between MSCs and cardiomyocytes. In HL-1 monolayers supplemented with MSC conditioned media (ConM) or tyrode (ConT) theta significantly increased in a time-dependent manner (ConT: t(0h): 2.4 cm/s +/- 0.2; t(4h): 3.1 +/- 0.4 cm/s), whereas the beating frequency remained constant. Connexin (Cx)43 mRNA and protein expression levels also increased after ConM or ConT treatment over the same time period. Enhanced low-density lipoprotein receptor-related protein 6 (LRP6) phosphorylation after ConT treatment implicates the Wnt signaling pathway. Suppression of Wnt secretion from MSCs (IWP-2; 5 mu mol/l) reduced the efficacy of ConT to induce phospho-LRP6 and to increase theta. Inhibition of beta-catenin (cardamonin; 10 mu mol/l) or GSK3-alpha/beta (LiCl; 5 mmol/l) also suppressed changes in theta, further supporting the hypothesis that MSC-mediated Cx43 upregulation occurs in part through secreted Wnt ligands and activation of the canonical Wnt signaling pathway.