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目的测定风湿性心脏瓣膜病(风心病)心房颤动(Af)心房肌缝隙连接蛋白(Connexin)40(Cx40)和Connexin43(Cx43)mRNA表达水平的变化。方法11例风心病患者[Af8例,窦性心律3例;男4例,女7例;平均年龄(50.8±13.7)岁;平均Af时间(5.6±6.5)年]的右心耳心肌1小块,提取所有标本的总RNA,等量转移至尼龙膜上,用PCR合成的特异cDNA探针,标记上放射性同位素(α32PdCTP)后与尼龙膜杂交,然后将杂交膜与X光胶片行放射自显影,在灰度扫描仪下得到每例标本mRNA水平的灰度值,为了进一步矫正转膜时可能的定量不统一,将同1块杂交膜先后与Cx40探针、Cx43探针和内参照βactin杂交,以Cx40/βactin和Cx43/βactin的百分比作为统计变量进行分析。结果在Af与窦性心律标本中,Cx40和Cx43的mRNA表达量未发生明显改变(P>0.05)。结论在风心病Af患者中,未发现缝隙连接蛋白Cx40和Cx43的mRNA表达量明显改变。
Objective To investigate the changes of the expression of connexin 40 (Cx40) and Connexin43 (Cx43) mRNA in atrial fibrillation (AF) patients with rheumatic heart disease (rheumatic heart disease). Methods A total of 11 patients with rheumatic heart disease [Af 8, sinus rhythm 3, male 4, female 7, mean age 50.8 ± 13.7, mean Af 5.6 5.6] The total RNA was extracted from all samples and transferred to nylon membrane in equal volume. The specific cDNA probes synthesized by PCR were labeled with radioisotope (α32PdCTP) and hybridized with nylon membrane. Then the hybridization membrane and X-ray film were autoradiographed , The grayscale value of mRNA of each sample was obtained under the gray scale scanner. In order to further correct the possible quantitative unification of the transfer membrane, the same hybrid membrane was crossed with Cx40 probe, Cx43 probe and internal reference βactin , Analyzed as a percentage of Cx40 / βactin and Cx43 / βactin as statistical variables. Results There was no significant change in mRNA expression of Cx40 and Cx43 in Af and sinus rhythm samples (P> 0.05). Conclusions In the Af patients with rheumatic heart disease, no significant changes in mRNA expression of Cx40 and Cx43 were observed.