Liu Yahua, Shen Hong, Wang Lixiang, Yang Huining, Ma Lizhi
Department of Emergency, General Hospital of Chinese People Armed Police Forces, Beijing 100039, China. Corresponding author: Shen Hong, Department of Emergency, General Hospital of PLA, Beijing 100853, China, Email:
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2015 Mar;27(3):209-12. doi: 10.3760/cma.j.issn.2095-4352.2015.03.010.
To investigate the effect of anisodamine on the expression of connexin 43 (Cx43) in swine ventricular myocardium after resuscitation from cardiac arrest.
The experiment was conducted on healthy pigs, and they were randomly divided into three groups, namely sham group, epinephrine group (control group) and anisodamine group (experimental group, animals were resuscitated combined with injection of 0.4 mg/kg of anisodamine), with 5 pigs in each group. Model of ventricular fibrillation was reproduced by alternating current challenge, and cardiopulmonary resuscitation (CPR) was performed 8 minutes after cardiac arrest. Left ventricular myocardium was harvested at 24 hours after restoration of spontaneous circulation. The expression and distribution of Cx43 were observed by immunofluorescence, Cx43 mRNA expression was assessed with reverse transcription-polymerase chain reaction (RT-PCR), and the protein expressions of Cx43 and phosphorylation of Cx43 (p-Cx43) were analyzed by Western Blot.
The positive expression of Cx43 in ventricular muscle was distributed uniformly, mostly at the end-to-end linkage of myocardial cells, with a few side-to-side linkage in sham group. The positive expression of Cx43 in control group was significantly weaker than that in the sham group, and the signal intensity was significantly declined (4.35±2.10 vs. 10.02±3.66, P < 0.01). The positive expression of Cx43 at the end-to-end linkage and side-to-side linkage was irregular in experimental group, and the signal intensity was obviously higher than that in the control group (7.91±2.54 vs. 4.35±2.10, P < 0.05), but it was significantly weaker than that in the sham group (7.91±2.54 vs. 10.02±3.66, P < 0.05). For control group and experimental group, the Cx43 mRNA and protein expressions were significantly lower than those of the sham group [Cx43 mRNA (A value): 0.32±0.05, 0.32±0.03 vs. 0.48±0.07; Cx43 protein (A value): 0.43±0.03, 0.50±0.07 vs. 0.65±0.04, all P < 0.01], and there were no significant differences between experimental group and control group (all P > 0.05). The p-Cx43 protein expression of control group was significantly lower than that of the sham group (A value: 0.22±0.03 vs. 0.37±0.06, P < 0.01), and it was significantly higher in the experiment group than that in the control group (A value: 0.29±0.07 vs. 0.22±0.03, P < 0.01), but there was no significant difference with the sham group (P > 0.05). No significant difference in p-Cx43/Cx43 was found among sham, control, and experiment groups (0.57±0.09, 0.51±0.05, 0.58±0.06, all P > 0.05).
Anisodamine can improve the abnormal expression of Cx43 in ventricular muscle of pigs with cardiac arrest, which may be related to the protection effect of anisodamine on cardiac conduction.
探讨山莨菪碱对猪心脏骤停复苏后心室肌中连接蛋白43（Cx43）表达的影响。
选用健康猪进行实验，随机分为三组，即假手术组、肾上腺素组（对照组）和山莨菪碱组（实验组，复苏时动物联合注射0.4mg/kg山莨菪碱），每组5只猪。通过交流电刺激复制心室颤动模型，心脏骤停8分钟后进行心肺复苏（CPR）。自主循环恢复24小时后采集左心室心肌。采用免疫荧光法观察Cx43的表达及分布，逆转录-聚合酶链反应（RT-PCR）检测Cx43 mRNA表达，蛋白质免疫印迹法分析Cx43蛋白表达及Cx43磷酸化（p-Cx43）情况。
假手术组心室肌Cx43阳性表达均匀分布，多位于心肌细胞端端连接，少量为侧侧连接。对照组Cx43阳性表达明显弱于假手术组，信号强度显著下降（4.35±2.10 vs. 10.02±3.66，P<0.01）。实验组端端连接和侧侧连接处Cx43阳性表达不规则，信号强度明显高于对照组（7.91±2.54 vs. 4.35±2.10，P<0.05），但显著弱于假手术组（7.91±2.54 vs. 10.02±3.66，P<0.05）。对照组和实验组Cx43 mRNA及蛋白表达均显著低于假手术组[Cx43 mRNA（A值）：0.32±0.05，0.32±0.03 vs. 0.48±0.07；Cx43蛋白（A值）：0.43±0.03，0.50±0.07 vs. 0.65±0.04，均P<0.01]，实验组与对照组比较差异无统计学意义（均P>0.05）。对照组p-Cx43蛋白表达显著低于假手术组（A值：0.22±0.03 vs. 0.37±0.06，P<0.01），实验组显著高于对照组（A值：0.29±0.07 vs. 0.22±0.03，P<0.01），但与假手术组比较差异无统计学意义（P>0.05）。假手术组、对照组和实验组p-Cx43/Cx43比较差异无统计学意义（0.57±0.09，0.51±0.05，0.58±0.06，均P>0.05）。
山莨菪碱可改善心脏骤停猪心室肌Cx43异常表达，这可能与山莨菪碱对心脏传导的保护作用有关。
