本文選題：地氟烷 + 單磷酸腺苷激活的蛋白激酶(AMPK)��； 參考：《中國老年學(xué)雜志》2017年04期

【摘要】：目的探討地氟烷預(yù)處理在大鼠離體心臟缺血再灌注(IR)中的保護(hù)作用是否與單磷酸腺苷激活的蛋白激酶(AMPK)有關(guān)。方法清潔級健康雄性大鼠50只,2~3月齡,體重250~280 g,切取離體心臟并隨機(jī)分為正常對照(C)組、IR組、地氟烷預(yù)處理+IR(DR)組、地氟烷預(yù)處理+AMPK抑制劑+IR(DC)組和IR+AMPK抑制劑(CR)組各10只。DR組和DC組在給予大鼠吸入1 MAC的地氟烷30 min后立即切取心臟。采用Langendorff灌注裝置并以95%O_2~5%CO_2飽和的K-H液作為灌注液,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。分別在停止灌注前和再灌注60 min時(shí)記錄心率(HR)、左室發(fā)展壓(LVDP)、左室內(nèi)壓最大上升速率(+dp/dt)和左室內(nèi)壓最大下降速率(-dp/dt)。采用caspase-3活性檢測試劑盒測定心肌組織內(nèi)caspase-3活性,酶聯(lián)免疫吸附(ELISA)法檢測冠脈流出液中心肌肌鈣蛋白(c Tn)I濃度。采用免疫組織化學(xué)法和Western印跡法檢測心肌組織內(nèi)AMPK(Thr-172)的磷酸化水平。結(jié)果與C組比較,IR組離體心臟再灌注60 min時(shí)HR、LVDP、+dp/dt和-dp/dt顯著降低(P0.05),心肌組織caspase-3活性和冠脈流出液中cTnI濃度顯著升高(P0.05),而AMPK(Thr-172)的磷酸化水平無顯著差異(P0.05)。與IR組比較,DR組HR、LVDP、+dp/dt和-dp/dt顯著升高(P0.05),caspase-3活性和cTnI濃度顯著降低(P0.05),AMPK(Thr-172)的磷酸化水平顯著升高(P0.05)。而與DR組比較,DC組HR、LVDP、+dp/dt和-dp/dt顯著降低(P0.05),caspase-3活性和cTnI濃度顯著升高(P0.05),AMPK(Thr-172)的磷酸化水平顯著降低(P0.05)。結(jié)論地氟烷預(yù)處理使IR后心肌組織內(nèi)AMPK(Thr-172)磷酸化水平升高而被激活,是其緩解心肌IR損傷的機(jī)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.
【作者單位】： 重慶醫(yī)科大學(xué)附屬第一醫(yī)院麻醉科;
【分類號】：R614

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