Department of Anesthesiology, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China.
J Zhejiang Univ Sci B. 2010 Sep;11(9):661-72. doi: 10.1631/jzus.B1000155.
Sevoflurane postconditioning reduces myocardial infarct size. The objective of this study was to examine the role of the phosphatidylinositol-3-kinase (PI3K)/Akt pathway in anesthetic postconditioning and to determine whether PI3K/Akt signaling modulates the expression of pro- and antiapoptotic proteins in sevoflurane postconditioning. Isolated and perfused rat hearts were prepared first, and then randomly assigned to the following groups: Sham-operation (Sham), ischemia/reperfusion (Con), sevoflurane postconditioning (SPC), Sham plus 100 nmol/L wortmannin (Sham+Wort), Con+Wort, SPC+Wort, and Con+dimethylsulphoxide (DMSO). Sevoflurane postconditioning was induced by administration of sevoflurane (2.5%, v/v) for 10 min from the onset of reperfusion. Left ventricular developed pressure (LVDP), left ventricular end-diastolic pressure (LVEDP), maximum increase in rate of LVDP (+dP/dt), maximum decrease in rate of LVDP (-dP/dt), heart rate (HR), and coronary flow (CF) were measured at baseline, R30 min (30 min of reperfusion), R60 min, R90 min, and R120 min. Creatine kinase (CK) and lactate dehydrogenase (LDH) were measured after 5 min and 10 min reperfusion. Infarct size was determined by triphenyltetrazolium chloride staining at the end of reperfusion. Total Akt and phosphorylated Akt (phospho-Akt), Bax, Bcl-2, Bad, and phospho-Bad were determined by Western blot analysis. Analysis of variance (ANOVA) and Student-Newman-Keuls' test were used to investigate the significance of differences between groups. The LVDP, + or - dP/dt, and CF were higher and LVEDP was lower in the SPC group than in the Con group at all points of reperfusion (P<0.05). The SPC group had significantly reduced CK and LDH release and decreased infarct size compared with the Con group [(22.9 + or - 8)% vs. (42.4 + or - 9.4)%, respectively; P<0.05]. The SPC group also had increased the expression of phospho-Akt, Bcl-2, and phospho-Bad, and decreased the expression of Bax. Wortmannin abolished the cardioprotection of sevoflurane postconditioning. Sevoflurane postconditioning may protect the isolated rat heart. Activation of PI3K and modulation of the expression of pro- and antiapoptotic proteins may play an important role in sevoflurane-induced myocardial protection.
七氟醚后处理可减少心肌梗死面积。本研究的目的是探讨磷脂酰肌醇 3-激酶(PI3K)/Akt 通路在麻醉后处理中的作用,并确定 PI3K/Akt 信号是否调节七氟醚后处理中促凋亡和抗凋亡蛋白的表达。首先制备分离和灌注的大鼠心脏,然后将其随机分配到以下组:假手术(Sham)、缺血/再灌注(Con)、七氟醚后处理(SPC)、Sham+100nmol/Lwortmannin(Sham+Wort)、Con+Wort、SPC+Wort 和 Con+二甲亚砜(DMSO)。七氟醚后处理通过在再灌注开始时给予七氟醚(2.5%,v/v)10 分钟来诱导。在基线、R30min(再灌注 30min)、R60min、R90min 和 R120min 时测量左心室发展压(LVDP)、左心室舒张末期压(LVEDP)、LVDP 的最大增加率(+dP/dt)、LVDP 的最大降低率(-dP/dt)、心率(HR)和冠状动脉流量(CF)。再灌注后 5min 和 10min 测量肌酸激酶(CK)和乳酸脱氢酶(LDH)。再灌注结束时通过氯化三苯基四氮唑染色确定梗死面积。通过 Western blot 分析测定总 Akt 和磷酸化 Akt(磷酸化 Akt)、Bax、Bcl-2、Bad 和磷酸化 Bad。使用方差分析(ANOVA)和 Student-Newman-Keuls 检验来研究组间差异的显著性。与 Con 组相比,再灌注各时点 SPC 组的 LVDP、+或-dP/dt 和 CF 更高,LVEDP 更低(P<0.05)。与 Con 组相比,SPC 组 CK 和 LDH 释放减少,梗死面积减小[(22.9+/-8)%比(42.4+/-9.4)%,分别;P<0.05]。SPC 组还增加了磷酸化 Akt、Bcl-2 和磷酸化 Bad 的表达,减少了 Bax 的表达。wortmannin 消除了七氟醚后处理的心脏保护作用。七氟醚后处理可能对分离的大鼠心脏具有保护作用。PI3K 的激活和促凋亡和抗凋亡蛋白表达的调节可能在七氟醚诱导的心肌保护中发挥重要作用。