家兔心房肌细胞环-磷酸腺苷动态变化的实时评价  

Real-time evaluation of the dynamic changes of cyclic adenosine monophosphate in atrial muscle cells of rabbits

作　　者：刘丽萍[1] 梁哲龙[1] 金正元[1] 金元哲[1] 崔勋[1] 

机构地区：[1]延边大学医学部生理学与病理生理学教研部,吉林省延吉市133000

出　　处：《中国临床康复》2006年第48期83-85,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金(30260033)~~

摘　　要：目的:探索实时评价心房肌细胞内环-磷酸腺苷动态变化的方法,为细胞内第二信使代谢水平分析提供可靠的实验数据。方法:实验于2004-04/2005-12在吉林省延边大学基础医学院生理学与病理生理学教研部完成。①采用大耳白家兔32只,建立跳动心房灌流模型,跳动心房稳定60min后,4℃下每2min收集灌流液样本。②采用固定心房跳动频率(1.3Hz)的方法,每2min收集1个灌流液样本,12min为1个循环。在1个对照循环之后,处理垂体腺苷酸环化酶激活肽(100nmol/L)或在非选择性磷酸酯酶抑制剂(1.0mmol/L)存在下再处理垂体腺苷酸环化酶激活肽1个循环,观察心房组织环-磷酸腺苷含量与逸出量的关系。③1个循环对照之后,给予30nmol/L的垂体腺苷酸环化酶激活肽或不同浓度的垂体腺苷酸环化酶激活肽(1,10,100nmol/L,每种浓度之间穿插2个循环的恢复期),实时观察心房肌细胞环-磷酸腺苷动态变化的特点。结果:①垂体腺苷酸环化酶激活肽对心房组织生成和逸出环-磷酸腺苷的影响:用垂体腺苷酸环化酶激活肽处理12min后,环-磷酸腺苷含量在组织和灌流液中分别增加到(42.18±11.17)nmol/g和(204.17±44.50)fkat/g,与对照值相比差异明显(P<0.05)。当给与非选择性磷酸酯酶抑制剂+垂体腺苷酸环化酶激活肽后,环-磷酸腺苷含量在组织和灌流液中分别增加到(149.90±13.87)nmol/g和(747.00±65.83)fkat/g(P<0.01)。且灌流液环-磷酸腺苷逸出量与组织环-磷酸腺苷含量之间存在线性关系(Y=0.2918X-0.118,R2=0.8809;P<0.01)。②垂体腺苷酸环化酶激活肽促进心房环-磷酸腺苷逸出量的情况:随着垂体腺苷酸环化酶激活肽浓度的增加,环-磷酸腺苷逸出量也随之增多,10,100nmol/L垂体腺苷酸环化酶激活肽的逸出峰值分别为(247.83±50.83),(254.00±53.50)fkat/g,均显著高于对照值和1nmol/L垂体腺苷酸环化酶激活肽[(66.00±7.50),(75.50±7.50)fkat/g;P<0.01],呈时间、剂量依AIM： To probe into real-time evaluating method of dynamic changes of adenosine phosphate, the cyclic adenosine monophosphate of atrial muscle cells, so as to provide reliable experimental data for analysis of metabolic level of second massenger inside the cells. METHODS： The experiment was conducted in the Department of Physiology and Pathophysiology, School of Basic Medicine of Yanbian University from April 2004 to December 2005. ①Thirty-two white rabbits were selected to establish beating atrial models. At 60 minutes after the beating atria were stabilized, the perfusate was collected at 4 ℃ with 2-minute intervals.②To analyze the relationship between Cyclic adenosine monophosphate （cAMP） efflux in the perfusate and the atrial cAMP content, the atria was paced at 1.3 Hz and the control period （12 minutes as an experimental cycle） was followed by an infusion of pituitary adenylate cyclase activating polypeptide （PACAP27, 100 nmol/L） or 3-isobutyl-1-methylxanthine （IBMX, 1.0 mmol/L）, an inhibitor of phosphodiesterases, plus PACAP27.③To analyze the effects of PACAP27 on cAMP production in real-time base, the control period was followed by an infusion of 30nmol/L and/or variable doses （1, 10, 100 nmol/L） of PACAP27 for 1 cycle, spaced by 2 cycles of recovery periods. RESULTS： ①PACAP27-induced content of cAMP in the atrial tissue and efflux of cAMP in the atrial perfusate were significantly increased to （42.18±11.17） nmol/g and （204.17±44.50） fkat/g, which was remarkably different with the control value （P 〈 0.05）. IBMX±PACAP27 also increased the atrial content and efflux of cAMP significantly [cAMP content： （149.90±13.87） nmol/L, cAMP efflux： （747.00±65.83） fkat/g; P 〈 0.01, vs. control value].The present study showing a linear correlation between cAMP efflux and cAMP production in beating rabbit atria （Y=0.291 8X-0.118, R^2=0.880 9; P 〈 0.01）. ②PACAP 27 increased cAMP production in a dose and timedependent manner with a minor effect on at

关 键 词：心房 环AMP 激肽类 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学]