地氟烷预处理对大鼠离体心脏缺血再灌注损伤的影响及作用机制

发布时间：2018-05-14 15:19

本文选题：地氟烷 + 单磷酸腺苷激活的蛋白激酶(AMPK)　；参考：《中国老年学杂志》2017年04期

【摘要】：目的探讨地氟烷预处理在大鼠离体心脏缺血再灌注(IR)中的保护作用是否与单磷酸腺苷激活的蛋白激酶(AMPK)有关。方法清洁级健康雄性大鼠50只,2~3月龄,体重250~280 g,切取离体心脏并随机分为正常对照(C)组、IR组、地氟烷预处理+IR(DR)组、地氟烷预处理+AMPK抑制剂+IR(DC)组和IR+AMPK抑制剂(CR)组各10只。DR组和DC组在给予大鼠吸入1 MAC的地氟烷30 min后立即切取心脏。采用Langendorff灌注装置并以95%O_2~5%CO_2饱和的K-H液作为灌注液,在平衡灌注15 min后,停止灌注30 min后再灌注60 min建立IR模型。分别在停止灌注前和再灌注60 min时记录心率(HR)、左室发展压(LVDP)、左室内压最大上升速率(+dp/dt)和左室内压最大下降速率(-dp/dt)。采用caspase-3活性检测试剂盒测定心肌组织内caspase-3活性,酶联免疫吸附(ELISA)法检测冠脉流出液中心肌肌钙蛋白(c Tn)I浓度。采用免疫组织化学法和Western印迹法检测心肌组织内AMPK(Thr-172)的磷酸化水平。结果与C组比较,IR组离体心脏再灌注60 min时HR、LVDP、+dp/dt和-dp/dt显著降低(P0.05),心肌组织caspase-3活性和冠脉流出液中cTnI浓度显著升高(P0.05),而AMPK(Thr-172)的磷酸化水平无显著差异(P0.05)。与IR组比较,DR组HR、LVDP、+dp/dt和-dp/dt显著升高(P0.05),caspase-3活性和cTnI浓度显著降低(P0.05),AMPK(Thr-172)的磷酸化水平显著升高(P0.05)。而与DR组比较,DC组HR、LVDP、+dp/dt和-dp/dt显著降低(P0.05),caspase-3活性和cTnI浓度显著升高(P0.05),AMPK(Thr-172)的磷酸化水平显著降低(P0.05)。结论地氟烷预处理使IR后心肌组织内AMPK(Thr-172)磷酸化水平升高而被激活,是其缓解心肌IR损伤的机制之一。
[Abstract]:Objective to investigate whether desflurane preconditioning may be related to adenosine monophosphate activated protein kinase (AMPK) in isolated rat heart ischemia reperfusion (IRI). Methods 50 healthy male rats of clean grade were divided into IR group and desflurane pretreatment group, which were pretreated with desflurane for 3 months, weighing 250 ~ 280g, and were divided randomly into two groups: normal control group (C) and desflurane pretreated group. Desflurane pretreated with AMPK inhibitor IRD (n = 10) and IR AMPK inhibitor CR-R (n = 10) and DC group (n = 10) were treated with desflurane for 30 min and the heart was removed immediately after inhalation of desflurane for 1 MAC. The IR model was established by using Langendorff perfusion device and 95%O_2~5%CO_2 saturated K-H solution as perfusion solution. After 15 min of equilibrium perfusion, 30 min after reperfusion and 60 min after reperfusion, the model of IR was established. Heart rate (HR), left ventricular development pressure (LVDPN), left ventricular pressure (DP / DT) and left ventricular pressure (LV / DP / DT) were recorded before and 60 min after reperfusion, respectively. Caspase-3 activity assay kit was used to detect caspase-3 activity in myocardial tissue, and enzyme linked immunosorbent assay (Elisa) was used to detect cardiac troponin C Tn)I concentration in coronary effluents. The phosphorylation level of AMPKG Thr-172 in myocardium was detected by immunohistochemistry and Western blotting. Results compared with group C, HRP, dp/dt and -DP / DT decreased significantly at 60 min after reperfusion. The activity of caspase-3 in myocardial tissue and the concentration of cTnI in coronary effluents increased significantly (P 0.05), but there was no significant difference in phosphorylation level of AMPK min Thr-172). Compared with IR group, HRP, dp/dt and -dp / DT in Dr group increased the activity of P0.05Caspase-3 and the concentration of cTnI significantly decreased the phosphorylation level of P0.05AMPKPr-172). Compared with Dr group, HRD, dp/dt and -dp / DT significantly decreased the activity of P0.05, and the concentration of cTnI significantly increased the phosphorylation level of P0.05AMPK-172in DC group. Conclusion desflurane preconditioning can increase the level of AMPKG Thr-172) phosphorylation in myocardium after IR, which is one of the mechanisms to alleviate myocardial IR injury.
【作者单位】：重庆医科大学附属第一医院麻醉科;
【分类号】：R614

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