国际麻醉学与复苏杂志   2019, Issue (1): 0-0
    
内源性孤啡肽通过对缝隙连接蛋白43的调节影响 大鼠急性心肌缺血时心律失常的发生
杨光, 熊畅, 韩毅, 杨建新1()
1.山西医科大学第二医院
Endogenous nociceptin/orphanin FQ affects the occurrence of arrhythmia in acute myocardial ischemia in rats through the regulation of connexin 43 protein
 全文:
摘要:

目的 观察大鼠急性心肌缺血引起的各类室性心律失常的发生和心肌缝隙连接蛋白43(connexin 43, Cx43)含量的变化以及孤啡肽(nociceptin/orphanin FQ, OFQ)受体拮抗剂UFP?蛳101对心律失常和Cx43的影响。 方法 雄性SD大鼠64只,体重200~250 g, 按照随机数字表法分为对照组(C组,24只,分为3个亚组,即冠状动脉结扎后15 min亚组、冠状动脉结扎后60 min亚组、冠状动脉结扎后120 min亚组,每组8只)、OFQ受体拮抗剂组(U组,24只,分为3个亚组,即冠状动脉结扎后15 min亚组、冠状动脉结扎后60 min亚组、冠状动脉结扎后120 min亚组,每组8只)、假手术组(S组,16只,分为两个亚组,即冠状动脉只穿线不结扎15 min亚组、冠状动脉只穿线不结扎60 min亚组,每组8只)。U组大鼠经尾静脉注射OFQ受体拮抗剂(UFP?蛳101,10-9 mol/L)1 μl/g,C组大鼠经尾静脉注射等体积的生理盐水溶液。C组、U组大鼠在给药5 min后结扎冠状动脉,S组冠状动脉只穿线不结扎。分别记录C组、U组大鼠从给药前10 min到冠状动脉结扎后15、60、120 min的ECG数据,分析统计各类室性心律失常。采用Western blot法测定各组标本中磷酸化Cx43(phosphorylate-Cx43, p-Cx43)以及总Cx43的含量。 结果 冠状动脉结扎后15 min时,与C组比较,U组室性期前收缩发生次数、联律的发生次数以及室性心动过速和心室纤颤(室颤)的持续时间明显下降(P<0.05)。冠状动脉结扎后60 min时,与C组比较,U组室性期前收缩发生次数、联律的发生次数以及室性心动过速和室颤的发生次数明显下降(P<0.05)。各组大鼠15 min亚组结果:与S组比较,C组p-Cx43表达明显减少(P<0.05);与C组比较,U组p-Cx43表达明显增加(P<0.05)。各组大鼠60 min 亚组结果:与S组比较,C组与U组p-Cx43表达均减少(P<0.05)。 结论 内源性OFQ拮抗剂UFP?蛳101通过促进Cx43蛋白发生磷酸化而减少大鼠在急性心肌缺血时心律失常的发生。

关键词: 内源性孤啡肽; 缝隙连接蛋白43; 磷酸化; 心律失常
Abstract:

Objective Observing influence of various ventricular arrhythmias induced by acute myocardial ischemia and the change of connexin 43 (Cx43) protein content in myocardium in rats and effects of nociceptin/orphanin FQ (OFQ) receptor antagonist UFP-101 on the expression of arrhythmia and Cx43 protein. Methods Sixty-four male SD rats , weighing 200-250 g, were randomly divided into control group(group C, each group was divided into three subgroups, which named coronary artery occlusion 15 min, coronary artery occlusion 60 min and coronary artery occlusion 120 min, n=8) , N/OFQ receptor antagonist group (group U, each group was divided into three subgroups, which named coronary artery occlusion 15 min, coronary artery occlusion 60 min and coronary artery occlusion 120 min, n=8), Sham operation group (group S, each group was divided into two subgroups, which named thread without ligation 15 min and thread without ligation 60 min, n=8). Group U was treated with OFQ antagonist compound UFP-101 in the dosage of 1 μl/g by caudal vein. Group C was treated with equal volume of saline. Group C and group U were administered 5 min before they were subjected to the coronary artery occlusion. Group S only was thread without ligation. Record data of electrocardiogram of group C and group U that from 10 min before the injection to 15, 60 min and 120 min after coronary artery occlusion, analyze data of all kinds of ventricular arrhythmia. Determination of expression levels of phosphorylated Cx43 (p-Cx43) protein and total Cx43 protein in all specimens by Western blot. Results At 15 min, numbers of ventricular premature contraction and ventricular bigeminy or trigeminy, the duration of ventricular tachycardia and ventricular fibrillation of group U were obviously lower than numbers in group C (P<0.05). At 60 min, numbers of ventricular premature contraction, ventricular bigeminy and trigeminy, ventricular tachycardia and ventricular fibrillation of group U were lower than those numbers in group C obviously (P<0.05). Expression of p-Cx43 protein at 15 min after coronary artery occlusion, the expression of p-Cx43 in group C were significantly lower than expression levels in group S (P<0.05), the expression of p-Cx43 in group U was significantly higher than that in group C (P<0.05). At 60 min after coronary artery occlusion, the expression of p-Cx43 in group C and group U were significantly lower than expression levels in group S (P<0.05). Conclusions Endogenous OFQ antagonist UFP-101 can reduce the occurrence of arrhythmia in acute myocardial ischemia by promoting phosphorylation of Cx43 protein in rats.

Key words: Endogenous nociceptin/orphanin FQ; Protein Cx43; Phosphorylation; Arrhythmia