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选择性阻断cGMP依赖性蛋白激酶对盲肠结扎穿孔致脓毒症大鼠血管环收缩功能的影响
Effects of selective blockade of cGMP-dependent protein kinase on contractility of vascular rings in a rat model of cecal ligation and puncture-induced sepsis.

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DOI:
作者:
王海燕，郭君，王学敏
WANG Haiyan，GUO Jun，WANG Xuemin
作者单位:
上海交通大学医学院附属第六人民医院麻醉科
Department of Anesthesiology，Shanghai Sixth People’s Hospital，Shanghai Jiaotong University of Medicine
关键词:
脓毒症；血管低反应性；血管环；cGMP依赖性蛋白激酶
Sepsis；Vascular hyporesponsiveness；Vascular ring；cGMP-dependent protein kinase
摘要:
目的 探讨选择性阻断cGMP依赖性蛋白激酶（PKG）对脓毒性休克大鼠血管低反应性的影响。 方法 雄性Sprague-Dawley（SD）大鼠随机分为假手术（SHAM）组、模型（CLP）组、一氧化氮合成酶抑制剂处理（L-NAME）组和PKG抑制剂处理（DT-2） 组，每组20只，以盲肠结扎穿孔法建立脓毒症模型，每组分别于造模后2、4、6h各取5只大鼠，分离胸主动脉制成血管环，L-NAME组和DT-2组血管在检测前分别与L-NAME或DT-2孵育30min。观测血管环对苯肾上腺素（PE）的浓度梯度效应曲线、最大收缩力（Emax)及产生最大收缩力的半数有效浓度（EC50）。于造模后4h各组取5只大鼠分离胸主动脉，以光谱法定量检测其PKG活性。 结果 CLP组血管环对PE的反应性较SHAM组降低（p＜0.05），且随造模时间延长血管环低反应性加重。L-NAME可明显提高造模后2、4、6h动脉环的收缩功能，Emax和EC50与CLP组相比差异明显（p＜0.05）。DT-2在造模后4h提高血管环的收缩功能，增加Emax值，降低EC50，较CLP组差异有统计学意义，造模后2h和6h，DT-2组与CLP组血管环收缩的浓度效应曲线差异无统计学意义（p＞0.05）。造模后4h，CLP组PKG活性较SHAM组显著升高（P＜0.01）；L-NAME组PKG活性较CLP组显著降低（P＜0.01）；DT-2组PKG活性明显低于CLP组（P＜0.05）。 结论 选择性阻断cGMP依赖性蛋白激酶可部分恢复脓毒症大鼠血管环低反应性。
Objective To investigate the effects of selective blockade of cGMP-dependent protein kinase on vascular hyporesponsiveness in septic shock. Methods Male Sprague-Dawley rats were randomly divided into SHAM group（n=20），CLP group（n=20），NG-nitro-L-arginine methyl ester (L-NAME) group（n=20） and DT-2 trifluoroacetate salt （DT-2）group（n=20）. Septic shock was induced by cecal ligation and puncture（CLP）. 5 rats were sacrificed in each group at 2、4 and 6h post-CLP respectively and thoracic aortas were prepared to measure the contraction-response curve，maximal tension （Emax） and the concentration that provide 50% maximal response（EC50）to phenylephrine（PE）using organ chamber technique. The aorta rings were incubated with L-NAME （L-NAME group） or DT-2 （DT-2 group）for 30min before the measurement. At 4 h post-CLP，the activity of PKG of aortic rings in each group was quantitatively measured by a spectrographic method in each group. Results The contractibility of the thoracic aortic rings from CLP group were significantly lower than those from SHAM group at 2，4 and 6h post-CLP（P＜0.05）.L-NAME effectively improved the contractibility of the thoracic aortic rings at 2、4 and 6h post-CLP ，and the Emax and EC50 of L-NAME group were significantly different from those of CLP group （P＜0.05）. DT-2 increased the contractibility of vascular rings just at 4 h post-CLP（P＜0.05），and the Emax and EC50 of DT-2 group were significantly different from those of CLP group （P＜0.01）. At 4 h post-CLP，the activity of PKG in CLP group was significantly higher than that in SHAM group（P＜0.01）；The activity in both L-NAME and DT-2 group was significantly lower than that in CLP group（P＜0.01 and P＜0.05，respectively）. Conclusion Selectively blocking cGMP-dependent protein kinase can partly restore the hyporesponsiveness of thoracic aortic rings in septic rats.

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