Li Lijuan, Hessel Marleen, van der Valk Lizet, Bax Minka, van der Linden Irma, van der Laarse Arnoud
Department of Cardiology, C5-P, Leiden University Medical Centre, P.O. Box 9600, 2300 RC, Leiden, The Netherlands.
Pflugers Arch. 2004 May;448(2):146-52. doi: 10.1007/s00424-003-1236-3. Epub 2004 Feb 6.
Although the troponins are the serum proteins most frequently used nowadays to diagnose myocardial infarction, controversy continues about whether troponins are released later from infarcted myocardium than the cytoplasmic enzymes used previously, like lactate dehydrogenase (LDH). The present study compared the release kinetics of troponin-I (TnI) and LDH from necrotic cardiomyocytes in vitro. Cardiomyocytes prepared from neonatal rat ventricles were grown for 3 days. A total of 126 cultures were subjected to metabolic inhibition to induce cell necrosis. At various time intervals cells and media were collected for quantitative analysis of LDH activity and TnI concentration. Mean (+/-SD) LDH activity and TnI content of nine cultures at time t=0 were 2.07+/-0.30 U and 1.52+/-0.30 micro g per culture, respectively. Release of LDH from necrotic cardiomyocytes preceded release of TnI by about 60 min. The quantity of LDH released from the cultures after 210 min was 83.2+/-10.0%, whereas that of TnI released after 210 min was always less (33.8+/-22.2%). Cytochemical assessment of necrotic cardiomyocytes showed TnI-positive cells that were poor in LDH. The delay of TnI release relative to LDH release may be explained by slow dissociation of TnI molecules from myofilaments and/or formation of TnI degradation products that are undetected by the currently used ELISA assay.
尽管肌钙蛋白是目前最常用于诊断心肌梗死的血清蛋白,但对于肌钙蛋白是否比以前使用的细胞质酶(如乳酸脱氢酶,LDH)从梗死心肌中释放得更晚,仍存在争议。本研究比较了肌钙蛋白I(TnI)和LDH在体外从坏死心肌细胞中的释放动力学。从新生大鼠心室制备的心肌细胞培养3天。总共126个培养物受到代谢抑制以诱导细胞坏死。在不同时间间隔收集细胞和培养基,用于定量分析LDH活性和TnI浓度。在t = 0时,九个培养物的平均(±标准差)LDH活性和TnI含量分别为每培养物2.07±0.30 U和1.52±0.30μg。坏死心肌细胞中LDH的释放比TnI的释放提前约60分钟。210分钟后从培养物中释放的LDH量为83.2±10.0%,而210分钟后释放的TnI量总是较少(33.8±22.2%)。坏死心肌细胞的细胞化学评估显示TnI阳性细胞中LDH含量较低。相对于LDH释放,TnI释放的延迟可能是由于TnI分子从肌丝上缓慢解离和/或形成了目前使用的ELISA检测未检测到的TnI降解产物。