IL-10 and TGF-β Increase Connexin-43 Expression and Membrane Potential of HL-1 Cardiomyocytes Coupled with RAW 264.7 Macrophages

Q3 Medicine
Cora B Cox, Mike Castro, T. Brown, N. J. Bigley
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引用次数: 1

Abstract

Cardiac resident macrophages facilitate electrical conduction by interacting with cardiomyocytes via connexin-43 (Cx43) hemichannels. Cx43 is critical for impulse propagation and coordination between muscle contractions. Cardiomyocyte electrophysiology can be altered when coupled with noncardiomyocyte cell types such as M2c tissue-resident macrophages. Using cocultures of murine HL-1 cardiomyocytes and RAW 264.7 macrophages, we examined the hypothesis that cytokine signals, TGF-β1 and IL-10, upregulate Cx43 expression at points of contact between the two cell types. These cytokine signals maintain the macrophages in an M2c anti-inflammatory phenotype, mimicking cardiac resident macrophages. The electrophysiology of cardiomyocytes was examined using di-8-ANEPPS potentiometric dye, which reflects a change in membrane potential. Greater fluorescence intensity of di-8-ANEPPS occurred in areas where macrophages interacted with cardiomyocytes. Suppressor of cytokine signaling 3 (SOCS3) peptide mimetic downregulated fluorescence of this membrane potentiometric stain. Cx43 expression in cocultures was confirmed by fluorescence microscopy and flow cytometry. Confocal images of these interactions demonstrate the Cx43 hemichannel linkages between the cardiomyocytes and macrophages. These results suggest that TGF-β1 and IL-10 upregulate Cx43 hemichannels, thus enhancing macrophage–cardiomyocyte coupling, raising the cellular resting membrane potential and leading to a more excitatory cardiomyocyte.
IL-10和TGF-β增加HL-1心肌细胞与RAW 264.7巨噬细胞偶联的连接素-43表达和膜电位
心脏巨噬细胞通过连接蛋白-43 (Cx43)半通道与心肌细胞相互作用,促进电传导。Cx43对冲动传播和肌肉收缩之间的协调至关重要。当与非心肌细胞类型(如M2c组织巨噬细胞)结合时,心肌细胞电生理可以改变。通过小鼠HL-1心肌细胞和RAW 264.7巨噬细胞共培养,我们检验了细胞因子信号TGF-β1和IL-10在两种细胞类型的接触点上调Cx43表达的假设。这些细胞因子信号维持巨噬细胞的M2c抗炎表型,模仿心脏巨噬细胞。采用反映膜电位变化的di8 - anepps电位计染料检测心肌细胞电生理。在巨噬细胞与心肌细胞相互作用的区域,di8 - anepps荧光强度更高。细胞因子信号传导3 (SOCS3)肽抑制因子模拟了这种膜电位染色的下调荧光。用荧光显微镜和流式细胞术证实Cx43在共培养中的表达。这些相互作用的共聚焦图像显示心肌细胞和巨噬细胞之间的Cx43半通道联系。这些结果表明,TGF-β1和IL-10上调Cx43半通道,从而增强巨噬细胞-心肌细胞偶联,提高细胞静息膜电位,使心肌细胞更兴奋。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
3.70
自引率
0.00%
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0
审稿时长
4 weeks
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