利用無特定病原豬建立缺血心肌保護機制之研究模式

The Establishment of SPF Pigs Model in Understanding Protective Mechanism of Myocardial Ischemia

林志鴻(Jyh-Hung Lin) ； 王耀宏(Yao-Horng Wang) ； 李文權(Wen-Chuan Lee) ； 李相台(Shiang-Tai Lee) ； 林凱元(Kai-Yuan Lin) ； 陳惠卿(Heuy-Chin Cheng) ； 林寶雪(Pao-Hsueh Lin) ； 黃琇琴(Hsiu-Chin Huang) ； 彭兆昌(Siu-Cheong Pang) ； 毛仁淡(Simon J. T. Mao) ； 楊平政(Ping-Cheng Yang) ； 朱瑞民(Redman Chu)

《中華民國獸醫學會雜誌》 24卷4期 (1998/12) Pp. 227-235

https://doi.org/10.7009/JTCSVS.199812.0227

摘要

Acute myocardial infarction may lead to myocardial ischemia and subsequent myocyte injury or even death. The synthesis of a 70 kDa heat shock protein (HSP70) induced by ischemic pretreatment have been considered to be one of the major causes to protect the ischemic myocardium against severe ischemic episode during the process of reperfusion. Recent evidences from transgenic mouse overexpressing human or rat inducible HSP70 have directly proved the cause-effect relationship between HSP synthesis and myocardial function. However, the regulatory mechanism controlling the expression of the HSP in the heart has not yet been well-examined. Since the extensive similarity of the cardiovascular system between pig and human. We, Pig Research Institute Taiwan (PRIT) with sophistic technology and dedicated facility, have chosen pig as an animal model to study the molecular mechanism of ischemic preconditioning. Nine specific pathogen free (SPF) pigs weighing about 30 kg were used. After tracheotomy, the left anterior decending coronary artery was gently dissected free of surrounding tissue after second branch and then subjected to a two cycles of 10 minutes occlusion and 30 minutes reperfusion . Three shamed-operated SPF pigs were served as controls. Electrocardiogram and the blood pressure of the pigs were monitored by polygraph showed that there were no significant difference between control and experimental animals. (p ＞ 0.05) Myocardium samples from infarction and non-infarction regions were collected and protein and mRNA levels for HSP70 were measured. The results from SDS-PAGE followed by immunoblotting showed no apparent difference in HSP70 levels. However, an inducible gene product of HSP70 was found to be increased to 2-10 folds when measured by using RT-PCR. The results indicate that porcine myocardium may augment gene expression of HSP70 in responding the ischemic damage. The successful establishment of this SPF pigs model will beneficial our understanding to the molecular mechanism of HSP to protect the ischemic myocardium.

關鍵字

缺血心肌保護機制 ； 豬模式 ； 熱緊迫蛋白 ； 無特定病原豬

並列摘要

Acute myocardial infarction may lead to myocardial ischemia and subsequent myocyte injury or even death. The synthesis of a 70 kDa heat shock protein (HSP70) induced by ischemic pretreatment have been considered to be one of the major causes to protect the ischemic myocardium against severe ischemic episode during the process of reperfusion. Recent evidences from transgenic mouse overexpressing human or rat inducible HSP70 have directly proved the cause-effect relationship between HSP synthesis and myocardial function. However, the regulatory mechanism controlling the expression of the HSP in the heart has not yet been well-examined. Since the extensive similarity of the cardiovascular system between pig and human. We, Pig Research Institute Taiwan (PRIT) with sophistic technology and dedicated facility, have chosen pig as an animal model to study the molecular mechanism of ischemic preconditioning. Nine specific pathogen free (SPF) pigs weighing about 30 kg were used. After tracheotomy, the left anterior decending coronary artery was gently dissected free of surrounding tissue after second branch and then subjected to a two cycles of 10 minutes occlusion and 30 minutes reperfusion . Three shamed-operated SPF pigs were served as controls. Electrocardiogram and the blood pressure of the pigs were monitored by polygraph showed that there were no significant difference between control and experimental animals. (p ＞ 0.05) Myocardium samples from infarction and non-infarction regions were collected and protein and mRNA levels for HSP70 were measured. The results from SDS-PAGE followed by immunoblotting showed no apparent difference in HSP70 levels. However, an inducible gene product of HSP70 was found to be increased to 2-10 folds when measured by using RT-PCR. The results indicate that porcine myocardium may augment gene expression of HSP70 in responding the ischemic damage. The successful establishment of this SPF pigs model will beneficial our understanding to the molecular mechanism of HSP to protect the ischemic myocardium.

並列關鍵字

Protective mechanism of myocardial ischemia ； Pig model ； Heat shock proteins ； Specific pathogen free (SPF) pigs