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目的研究正常窦律(SR)与心房颤动(AF)两组患者肺静脉前庭(PVA)组织中T型钙离子通道(T-type calcium channel,TTCC)α1G、α1H亚基mRNA及蛋白的表达差异。方法术中获取各组患者(SR=34,AF=38)少量肺静脉前庭组织标本,实时荧光定量RT-PCR对各组α1G及α1H亚基的mRNA丰度相对定量(2-ΔΔCt法);蛋白免疫印迹法比较两种α1亚基蛋白表达水平;HE染色及免疫组化染色比较两种α1亚基蛋白在细胞中的分布表达。结果与SR组相比,AF组患者的肺静脉前庭组织中TTCC的α1H亚基的mRNA表达丰度及亚基蛋白的表达水平均明显升高,但α1G亚基在两组中的表达水平接近,H-E染色及免疫组织化学染色结果与前一结论符合,并证实α1G、α1H亚基蛋白定位于心肌细胞膜表面。结论 AF组对比SR组,其T型钙通道α1H亚基表达显著增加,而α1G亚基的表达水平两组间无明显差异。
Objective To investigate the expression of α1G and α1H subunit mRNA and protein in T-type calcium channel (TTCC) in the vestibular veins of the normal sinus rhythm (SR) and atrial fibrillation (AF) patients. Methods A small amount of vestibular tissue samples of pulmonary veins were obtained during operation in each group (SR = 34, AF = 38). The mRNA abundance of α1G and α1H subunits in each group was determined by real-time fluorescence quantitative RT-PCR (2-ΔΔCt method) Western blotting was used to compare the expression of two α1 subunit proteins. HE staining and immunohistochemical staining were used to analyze the distribution of the two α1 subunits in the cells. Results Compared with SR group, the mRNA abundance of α1H subunit and the expression of subunit protein of TTCC in AF group were significantly increased in AF group, but the expression level of α1G subunit in both groups were close, HE staining and immunohistochemical staining results consistent with the previous conclusion, and confirmed α1G, α1H subunit protein localized in the myocardial cell membrane surface. Conclusion Compared with SR group, the expression of α1H subunit of T-type calcium channel in SR group is significantly increased, while the expression level of α1G subunit is not significantly different between two groups.