冷刺激对乳鼠心肌细胞缝隙连接蛋白43的影响及药物干预的研究  

Effect of Cold Stress on Connexin 43 Protein Expression With Drug Intervention in Neonatal Rats' Myocardial Cells

作　　者：黄强辉[1] 洪葵[1] 程晓曙[1] 胡建新[1] 

机构地区：[1]南昌大学第二附属医院心内科,江西省南昌市330006

出　　处：《中国循环杂志》2015年第1期59-63,共5页Chinese Circulation Journal

摘　　要：目的:在急性冷刺激乳鼠心肌细胞模型下,评价缝隙连接蛋白43(Connexin43,Cx43)的表达及研究急性冷刺激时乳鼠心肌细胞间传导的变化及其机制。方法 :乳鼠心肌细胞原代培养,随机分为正常对照组,实验4℃组;实验0℃组;并在实验4℃组和实验0℃组分别加入100 nmol/L抗心律失常肽(APP)10,每组乳鼠8只。采用流式细胞术(FCM)分析各组细胞的凋亡率,应用聚合酶链反应(PCR)和蛋白免疫印迹法(Western blot)检测Cx43及其磷酸化水平在基因和蛋白上的表达。结果 :FCM结果显示:实验4℃组和实验0℃组随着冷刺激程度加强和低温时间的延长,心肌细胞的凋亡率增加;同时PCR结果显示:实验4℃组和实验0℃组Cx43 m RNA呈现不同程度的下降;Western blot结果显示:实验4℃组和实验0℃组随着刺激强度和低温时间的延长,Cx43蛋白表达亦出现不同程度的下降;磷酸化的Cx43(P-Cx43)蛋白表达亦下降,而AAP10干预后可提高两组Cx43和P-Cx43蛋白的表达。结论 :心肌细胞在急性冷刺激时Cx43在基因和蛋白水平上表达均有不同程度的下降,P-Cx43表达亦下降,而AAP10能提高Cx43和P-Cx43蛋白的表达从而改善细胞间的传导。Objective： To study the effects of acute cold stress on connexin43 （Cx43） protein expression with drug intervention, and cell to cell conduction with its mechanism in neonatal rats’ myocardial cells. Methods： The primary neonatal rats’ myocardial cell culture was conducted in 4 groups. Group① , the cells were normally cultured, Group②, the cells were cultured at 4℃, Group③, the cells were cultured at 0℃and Group④, the anti-arrhythmia peptide （AAP 10） was added in Group②and Group③. The apoptosis rate of myocardial cells was evaluated by lfow cytometry assay, mRNA and protein expressions of CX43 were examined by RT-PCR and Western blot analysis, and CX43 phosphorylation product （P-CX43） was detected. Results： Compared with normally cultured cells, the myocardial cell apoptosis rate was obviously increased by acute cold stress at 4℃and 0℃with time extension. The mRNA expression of Cx43 was decreased at varying degrees at 4℃and 0℃stimulation, the protein expression of Cx43 was decreased at varying degrees at 4℃and 0℃stimulation with time extension, and P-Cx43 level was decreased. While the APP 10 intervention may obviously elevate the protein levels of Cx43 and P-Cx43. Conclusion： Acute cold stress could reduce the protein expression of CX43 and P-CX43, while APP 10 intervention may elevate such expression and improve the cell to cell conduction in neonatal rats’ myocardial cells.

关 键 词：冷刺激 心肌细胞 缝隙连接蛋白43 磷酸化 抗心律失常肽 CONNEXIN43 

分 类 号：R54[医药卫生—心血管疾病]