Kochan Z, Smolenski R T, Yacoub M H, Seymour A L
Department of Cardiothoracic Surgery, National Heart and Lung Institute at Harefield Hospital, UK.
J Mol Cell Cardiol. 1994 Nov;26(11):1497-503. doi: 10.1006/jmcc.1994.1168.
Evaluation of enzyme activities involved in nucleotide metabolism and adenosine production within different cell types can provide important information on their contribution to the overall metabolism of the heart. The following enzyme activities were determined: adenosine kinase (AK), adenosine deaminase (ADA), S-adenosylhomocysteine hydrolase (SAHH), purine nucleoside phosphorylase (PNP), AMP deaminase (AMPD), membrane 5'nucleotidase (M5'N), AMP specific (AC5'N) and IMP specific (IC5'N) cytosolic 5'nucleotidases in (1) rat heart (n = 5), (2) rat cardiomyocytes obtained by collagenase digestion (n = 5), (3) human heart (n = 6) obtained from explants or papillary muscles collected during heart transplantation or mitral valve replacement, and (4) human umbilical cord endothelial cells in primary culture (n = 4). In the human heart, activities (mumol/min/g wet weight) were as follows: AK (0.14 +/- 0.01), ADA (0.46 +/- 0.03), SAHH (0.001 +/- 0.0003), PNP (0.43 +/- 0.08), AMPD (0.41 +/- 0.05), M5'N (1.75 +/- 0.12), IC5'N (0.21 +/- 0.03) and AC5'N (0.11 +/- 0.02). These enzyme activities were lower than those determined in the rat heart with the exception of AC5'N and IC5'N which were equal. The most prominent difference observed was for AMPD and M5'N which were nine and five-fold more active in the rat heart. Rat cardiomyocyte enzyme activities were comparable to those measured in whole rat heart with the exception of ADA (six-fold lower) and PNP (16-fold lower). Endothelial cell activities were notably different from those in the human heart particularly in the case of SAHH (nine-fold higher) and PNP (16-fold higher).(ABSTRACT TRUNCATED AT 250 WORDS)
评估不同细胞类型中参与核苷酸代谢和腺苷生成的酶活性,可为它们对心脏整体代谢的贡献提供重要信息。测定了以下酶活性:腺苷激酶(AK)、腺苷脱氨酶(ADA)、S-腺苷同型半胱氨酸水解酶(SAHH)、嘌呤核苷磷酸化酶(PNP)、AMP脱氨酶(AMPD)、膜5'核苷酸酶(M5'N)、AMP特异性(AC5'N)和IMP特异性(IC5'N)胞质5'核苷酸酶,样本包括:(1)大鼠心脏(n = 5);(2)通过胶原酶消化获得的大鼠心肌细胞(n = 5);(3)从心脏移植或二尖瓣置换术中获取的外植体或乳头肌得到的人心脏(n = 6);以及(4)原代培养的人脐静脉内皮细胞(n = 4)。在人心脏中,酶活性(μmol/分钟/克湿重)如下:AK(0.14±0.01)、ADA(0.46±0.03)、SAHH(0.001±0.0003)、PNP(0.43±0.08)、AMPD(0.41±0.05)、M5'N(1.75±0.12)、IC5'N(0.21±0.03)和AC5'N(0.11±0.02)。除AC5'N和IC5'N活性相当外,这些酶活性均低于在大鼠心脏中测定的活性。观察到的最显著差异在于AMPD和M5'N,它们在大鼠心脏中的活性分别高出9倍和5倍。大鼠心肌细胞的酶活性与整个大鼠心脏中测得的活性相当,但ADA(低6倍)和PNP(低16倍)除外。内皮细胞的活性与人心肌细胞的活性明显不同,尤其是SAHH(高9倍)和PNP(高16倍)。(摘要截断于250字)