Yan Hui, Chen Jun-zhu, Zhu Jian-hua, Ni Yi-ming, Yu Guo-wei, Hu Shen-jiang, Tao Qian-min
Department of Cardiology, First Affiliated Hospital of Medical School of Zhejiang University, Hangzhou 310003, China.
Zhonghua Yi Xue Za Zhi. 2004 Feb 2;84(3):209-13.
To explore the expression of connexin (Cx) and its signal transduction pathway in the atrium of patients with atrial fibrillation (AF).
Atrial tissue samples of 63 patients undergoing cardiac surgery, including patients with chronic AF or paroxysmal AF (PAF), and those with sinus rhythm, were collected during operation. The mRNA expressions of calcineurin B and mitogen-activated protein kinase-1 (MKP-1) were detected by semi-quantitative RT-PCR. The protein expressions of extracellular-signal regulated kinase 1 (ERK1), phospho-ERK1 (P-ERK1), Cx40 and Cx43 were analyzed by Western blotting. HE staining and immunohistochemistry were used to examine the distribution of Cx40 and Cx 43.
Increased amounts of Cx40 protein (left atrium: 2.2 +/- 0.8, 2.2 +/- 0.6; right appendages: 2.1 +/- 0.5, 2.0 +/- 0.8) were found in the left atrium and right appendages of patients with Chronic persistent AF (CAF) or paroxysmal AF (PAF) (P < 0.05). The expression of Cx43 was only increased in the left atrium of patients with CAF and PAF (3.1 +/- 0.6, 2.8 +/- 0.7 vs 1.0 +/- 0.2, both P < 0.05). The amounts of Calcineurin B mRNA, MKP-1 mRNA and P-ERK1 of patients with CAF or PAF were significantly increased compared with patients in sinus rhythm (P < 0.05). Immunohistochemistry revealed that Cx40 and Cx43 of CAF patients and PAF patients acculmated in the intracellular site, and at the lateral member of atrial cells, both connexins redistributed.
The increased expression and disorderly distribution of Cx 40 and Cx 43 protein in the atrium of AF patients may be related with the abnormal activation and disequilibria of regulation of ERK1, MKP-1 and calcineurin.
探讨连接蛋白(Cx)及其信号转导通路在心房颤动(AF)患者心房中的表达情况。
收集63例接受心脏手术患者的心房组织样本,包括慢性房颤或阵发性房颤(PAF)患者以及窦性心律患者。术中采用半定量逆转录聚合酶链反应(RT-PCR)检测钙调神经磷酸酶B和丝裂原活化蛋白激酶-1(MKP-1)的mRNA表达。采用蛋白质印迹法分析细胞外信号调节激酶1(ERK1)、磷酸化ERK1(P-ERK1)、Cx40和Cx43的蛋白表达。采用苏木精-伊红(HE)染色和免疫组织化学法检测Cx40和Cx43的分布。
在慢性持续性房颤(CAF)或阵发性房颤(PAF)患者的左心房和右心耳中发现Cx40蛋白含量增加(左心房:2.2±0.8,2.2±0.6;右心耳:2.1±0.5,2.0±0.8)(P<0.05)。Cx43的表达仅在CAF和PAF患者的左心房中增加(3.1±0.6,2.8±0.7对比1.0±0.2,均P<0.05)。与窦性心律患者相比,CAF或PAF患者的钙调神经磷酸酶B mRNA、MKP-1 mRNA和P-ERK1含量显著增加(P<0.05)。免疫组织化学显示,CAF患者和PAF患者的Cx40和Cx43在细胞内位点积聚,并且在心房细胞的侧缘,两种连接蛋白均重新分布。
AF患者心房中Cx40和Cx43蛋白表达增加及分布紊乱可能与ERK1、MKP-1和钙调神经磷酸酶的异常激活和调节失衡有关。