Ding Chandong, Yang Cuicui, Cao Quanxia, Zhu Xiaoxia, Zhang Jianming, Zhang Wen, Wang Yongping, Li Long
Department of Cardiology, The Second Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230601, P.R. China.
Catheter Intervention Center, The Second Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230601, P.R. China.
Exp Ther Med. 2017 Sep;14(3):2108-2116. doi: 10.3892/etm.2017.4789. Epub 2017 Jul 12.
Currently, the mechanism of the chronotropic ability of stem cells modified to express the hyperpolarization-activated cyclic nucleotide-gated (HCN) gene remains to be elucidated. The present study assessed the effects of mouse (m)HCN2 gene modification on the expression of chronotropic relevant receptors, adrenergic receptor β1 (Adrb1) and cholinergic receptor muscarinic M2 (Chrm2), in bone marrow stromal cells (BMSCs) co-cultured with atrial myocytes. BMSCs were divided into the following four groups: i) BMSCs transfected with the mHCN2 gene and co-cultured with atrial myocytes for 48 h (TF + CO); ii) respective transfection (TF); iii) respective co-culture (CO); and iv) the control group without treatment (CTL). Green fluorescent protein (GFP) was observed in the BMSCs 48 h after transfection with pEGFP-C1-mHCN2. The expression of Adrb1 and Chrm2 was significantly increased in the TF and TF + CO groups, particularly the TF + CO group, compared with the CTL group (P<0.05). This suggests that BMSCs modified to express the mHCN2 gene possess autorhythmicity and chronotropic ability, particularly when co-cultured with atrial myocytes. The results of the present study provide novel information regarding the molecular basis of biological pacemakers' chronotropic ability.
目前，经修饰以表达超极化激活的环核苷酸门控（HCN）基因的干细胞变时能力机制仍有待阐明。本研究评估了小鼠（m）HCN2基因修饰对与心房肌细胞共培养的骨髓基质细胞（BMSC）中变时相关受体肾上腺素能受体β1（Adrb1）和胆碱能毒蕈碱型M2受体（Chrm2）表达的影响。BMSC分为以下四组：i）用mHCN2基因转染并与心房肌细胞共培养48小时的BMSC（TF + CO）；ii）各自转染（TF）；iii）各自共培养（CO）；iv）未处理的对照组（CTL）。用pEGFP-C1-mHCN2转染后48小时在BMSC中观察到绿色荧光蛋白（GFP）。与CTL组相比，TF组和TF + CO组中Adrb1和Chrm2的表达显著增加，尤其是TF + CO组（P<0.05）。这表明经修饰以表达mHCN2基因的BMSC具有自律性和变时能力，特别是与心房肌细胞共培养时。本研究结果提供了关于生物起搏器变时能力分子基础的新信息。
