Menzele Amanda, Aboalgasm Hamida, Ballo Robea, Gwanyanya Asfree
Department of Human Biology, Faculty of Health Sciences, University of Cape Town, Observatory, Cape Town, 7925, South Africa.
Histochem Cell Biol. 2023 Apr;159(4):329-337. doi: 10.1007/s00418-022-02170-5. Epub 2022 Dec 22.
Diabetes mellitus with hyperglycaemia is a major risk factor for malignant cardiac dysrhythmias. However, the underlying mechanisms remain unclear, especially during the embryonic developmental phase of the heart. This study investigated the effect of hyperglycaemia on the pulsatile activity of stem cell-derived cardiomyocytes. Mouse embryonic stem cells (mESCs) were differentiated into cardiac-like cells through embryoid body (EB) formation, in either baseline glucose or high glucose conditions. Action potentials (APs) were recorded using a voltage-sensitive fluorescent dye and gap junction activity was evaluated using scrape-loading lucifer yellow dye transfer assay. Molecular components were detected using immunocytochemistry and immunoblot analyses. High glucose decreased the spontaneous beating rate of EBs and shortened the duration of onset of quinidine-induced asystole. Furthermore, it altered AP amplitude, but not AP duration, and had no impact on neither the expression of the hyperpolarisation-activated cyclic nucleotide-gated isoform 4 (HCN4) channel nor on the EB beating rate response to ivabradine nor isoprenaline. High glucose also decreased both the intercellular spread of lucifer yellow within an EB and the expression of the cardiac gap junction protein connexin 43 as well as upregulated the expression of transforming growth factor beta 1 (TGF-β1) and phosphorylated Smad3. High glucose suppressed the autorhythmicity and gap junction conduction of mESC-derived cardiomyocytes, via mechanisms probably involving TGF-β1/Smad3 signalling. The results allude to glucotoxicity related proarrhythmic effects, with potential clinical implications in foetal diabetic cardiac disease.
伴有高血糖的糖尿病是恶性心律失常的主要危险因素。然而,其潜在机制仍不清楚,尤其是在心脏胚胎发育阶段。本研究调查了高血糖对干细胞衍生心肌细胞搏动活性的影响。将小鼠胚胎干细胞(mESCs)在基础葡萄糖或高葡萄糖条件下通过胚体(EB)形成分化为类心脏细胞。使用电压敏感荧光染料记录动作电位(APs),并使用刮擦加载荧光黄染料转移试验评估缝隙连接活性。使用免疫细胞化学和免疫印迹分析检测分子成分。高葡萄糖降低了EB的自发搏动率,并缩短了奎尼丁诱导的心搏停止发作的持续时间。此外,它改变了AP幅度,但未改变AP持续时间,并且对超极化激活的环核苷酸门控亚型4(HCN4)通道的表达以及EB对伊伐布雷定或异丙肾上腺素的搏动率反应均无影响。高葡萄糖还降低了荧光黄在EB内的细胞间扩散以及心脏缝隙连接蛋白连接蛋白43的表达,并上调了转化生长因子β1(TGF-β1)和磷酸化Smad3的表达。高葡萄糖通过可能涉及TGF-β1/Smad3信号传导的机制抑制了mESC衍生心肌细胞的自律性和缝隙连接传导。这些结果暗示了与糖毒性相关的促心律失常作用,对胎儿糖尿病性心脏病具有潜在的临床意义。