Interaction between cyclic GMP protein kinase and cyclic AMP may be diminished in stunned cardiac myocytes.
Interaction between cyclic GMP protein kinase and cyclic AMP may be diminished in stunned cardiac myocytes.
复制标题
在震惊的心肌细胞中，环 GMP 蛋白激酶和环 AMP 之间的相互作用可能会减弱。
DOI：
10.1016/s0014-2999(01)01216-x
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发表时间：
2001
影响因子：
5
通讯作者：
Weiss,HR
中科院分区：
文献类型：
--
作者：
Yan,L;Patel,KN;Zhang,Q;Scholz,PM;Weiss,HR
我们测试了一种假设，即在顿抑(模拟缺血/再灌流)的心肌细胞中，循环GMP蛋白激酶的负面功能效应的重要性将会降低。取新西兰大白兔(N=7)，分离出心室肌细胞。分别在基础状态和模拟缺血(37℃，95%N_2-5%CO_2 15分钟)和模拟再灌流(复氧)后观察心肌细胞的变化。用视频边缘检测器研究细胞缩短；用氧气电极测量氧气消耗。凝胶电泳后用放射自显影测定蛋白质的磷酸化。(1)8-(4-chlorophenylthio)guanosine-3′，5′-monophosphate(N2-etheno-8-bromoguanosine-3′，5′-monophosphorothioate，，cGMP蛋白激酶激动剂)10−7或10−5M；(2)8-BR-cAMP 10−5M后，PCPT 10−7或10−5M；(3)β-苯基-1，cGMP SP异构体(SP，cGMP蛋白激酶激动剂)10−7或10−5M；(2)8-BR-cAMP 10−5M，SP 10−7或10−5M。在基础状态下，顿抑心肌细胞的短轴缩短率(Pcs)和最大短轴缩短率(Rs)显著降低(Pcs：5.0±0.2%对3.8±0.3；Rs：64.8±5.9μm/S对46.9±4.8)。在两组中，PCPT和SP均呈剂量依赖性地降低PCs和Rs。对晕厥的心肌细胞的影响轻微，但并不显著。8-溴-环腺苷显著增加对照组的心肌细胞功能，但对顿抑心肌细胞无明显影响(pCS，4.5±0.5比6.2±0.8比3.1±0.2比3.6±0.2)。对照组经8-溴-环腺苷后，PCPT和SP的负性功能作用减弱(从−的39%降至−的29%)，而顿抑心肌细胞的负性作用更明显(−为19%)。PCPT和环状AMP具有相似的蛋白条带。在顿抑心肌细胞中，PCPT对三条(22、31和53 kDa)条带的增强作用较弱。
We tested the hypothesis that the importance of the negative functional effects of the cyclic GMP protein kinase would be reduced in stunned (simulated ischemia/reperfusion) cardiac myocytes. Ventricular cardiac myocytes were isolated from New Zealand white rabbits (N=7). Myocytes were studied at baseline and after simulated ischemia (15 min of 95% N2–5% CO2at 37 °C) followed by simulated reperfusion (reoxygenation). Cell shortening was studied with a video edge detector; O2consumption was measured using O2electrodes. Protein phosphorylation was measured autoradiographically after gel electrophoresis. Functional and metabolic data were acquired after: (1) 8-(4-chlorophenylthio)guanosine-3′,5′-monophosphate (PCPT, cGMP protein kinase agonist) 10−7or 10−5M; (2) 8-Br-cAMP 10−5M followed by PCPT 10−7or 10−5M; (3) β-phenyl-1, N2-etheno-8-bromoguanosine-3′,5′-monophosphorothioate, SP-isomer (SP, cGMP protein kinase agonist) 10−7or 10−5M; (2) 8-Br-cAMP 10−5M followed by SP 10−7or 10−5M. At baseline, percent of shortening (Pcs) and maximal rate of shortening (Rs) were significantly lower in the stunned myocytes (Pcs: 5.0±0.2% control vs. 3.8±0.3 stunned; Rs: 64.8±5.9 μm/s control vs. 46.9±4.8 stunned). In both groups, PCPT and SP dose-dependently decreased Pcs and Rs. The effects were slightly, but not significantly, less in stunned myocytes. 8-Br-cyclic AMP significantly increased function in control, but not stunned myocytes (Pcs, 4.5±0.5 to 6.2±0.8 control vs. 3.1±0.2 to 3.6±0.2 stunned). The negative functional effects of PCPT and SP were diminished after 8-Br-cyclic AMP in control (from −39% to−29%) and diminished significantly more in the stunned myocytes (−19%). PCPT and cyclic AMP phosphorylated similar protein bands. In stunned myocytes, three (22, 31 and 53 kDa) bands were enhanced less by PCPT.