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目的:研究急性缺氧时缝隙连接蛋白Cx43磷酸化水平的改变及抗心律失常肽(AAP)10对其影响。方法:大鼠体外心脏灌流模型,随机分为对照组、缺氧30min组、AAP10干预组,每组各9只。用Western-Blot技术检测Cx43磷酸化水平的改变,用免疫荧光双标结合激光扫描共聚焦成像技术显示Cx43分布的改变。结果:Western-Blot结果显示:缺氧30min组总Cx43蛋白表达下降(P<0.05),而去磷酸化的Cx43(NP-Cx43)蛋白表达不变。AAP10干预组能提高总Cx43蛋白表达(P<0.05),但对NP-Cx43蛋白表达无影响。而免疫荧光结果显示:缺氧30min组总Cx43和NP-Cx43蛋白分布均明显减少,且AAP10干预组仅能提高总Cx43蛋白表达,但对NP-Cx43蛋白无影响。结论:急性缺氧时细胞间通讯功能下降主要由于磷酸化的Cx43蛋白水平下降,而其中NP-Cx43蛋白的内化也可能参与该影响因素。而AAP10改善传导主要通过促进Cx43磷酸化而发挥作用。
AIM: To investigate the changes of Cx43 phosphorylation in acute hypoxia and the effect of anti-arrhythmic peptide (AAP) 10 on it. Methods: Rat heart perfusion model was randomly divided into control group, hypoxia 30min group, AAP10 intervention group, each group of 9 rats. The level of Cx43 phosphorylation was detected by Western-Blot and the distribution of Cx43 was detected by immunofluorescence double labeling combined with laser scanning confocal imaging. Results: The results of Western-Blot showed that the expression of total Cx43 protein decreased in hypoxia 30min group (P <0.05), while the dephosphorylated Cx43 (NP-Cx43) protein expression did not change. AAP10 intervention group can increase the total Cx43 protein expression (P <0.05), but had no effect on the expression of NP-Cx43 protein. The results of immunofluorescence showed that the distribution of total Cx43 and NP-Cx43 protein in hypoxia 30min group were significantly decreased, and the AAP10 intervention group only increased the total Cx43 protein expression, but had no effect on NP-Cx43 protein. CONCLUSION: The decrease of cell-cell communication function in acute hypoxia is mainly due to the decrease of phosphorylated Cx43 protein, and the internalization of NP-Cx43 protein may also be involved in the influencing factors. However, AAP10 improves transduction and plays a role mainly through promoting Cx43 phosphorylation.