摘要
目的从PI3K/Akt/mTOR通路角度,探讨七氟醚对小鼠心肌缺血再灌注损伤中自噬水平的影响。方法小鼠24只随机分4组(n=6):假手术组,只开胸不结扎;缺血再灌注组,戊巴比妥钠麻醉下结扎冠状动脉左前降支行缺血/再灌注手术;七氟醚组,七氟醚麻醉下手术;七氟醚+LY294002组,术前一周每日注射PI3K阻断剂LY294002,余同七氟醚组。收集血浆检测心肌损伤标志物肌酸激酶-MB(CK-MB)和心肌肌钙蛋白Ⅰ(cTnI)水平。心肌组织用免疫印迹法检测自噬标志物LC3、Beclin1,及信号通路磷脂酰肌醇3-磷酸激酶(PI3K)、蛋白激酶B(Akt)、雷帕霉素(mTOR)总蛋白及磷酸化蛋白水平。结果与假手术组相比,其余3组的血浆CK-MB、cTnI、心肌自噬标志蛋白LC3和Beclin1水平均增高(P<0.05),磷酸化PI3K、Akt、mTOR与总蛋白比值降低(P<0.05)。与假手术组相比,七氟醚组的PI3K、Akt、mTOR总蛋白水平差异无统计学意义;而缺血再灌注组PI3K、Akt水平增高(P<0.05),且mTOR蛋白水平降低(P<0.05);七氟醚+LY294002组PI3K水平增高(P<0.05),且Akt、mTOR蛋白水平降低(P<0.05)。与缺血再灌注组相比,七氟醚组和七氟醚+LY294002组血浆CK-MB和cTnI、LC3和Beclin1水平均降低(P<0.05)。与缺血再灌注组相比,七氟醚组的磷酸化PI3K、Akt、mTOR蛋白与总蛋白比值升高(P<0.05)。与七氟醚组相比,七氟醚+LY294002组血浆CK-MB、cTnI降低,LC3和Beclin1水平均增高(P<0.05);七氟醚+LY294002组的磷酸化PI3K、Akt、mTOR蛋白与总蛋白比值均降低(P<0.05)。结论七氟醚抑制了缺血再灌注引发的过度自噬,可能的机制是上调了PI3K/Akt/mTOR通路蛋白磷酸化水平。
Objective From the perspective of PI3 K/Akt/mTOR pathway,to explore the potential effect of sevoflurane on autophagy levels in mice with myocardial ischemia-reperfusion injury.Methods Twenty-four mice were randomly divided into 4 groups(n=6):sham operation group(only chest opening without ligation),ischemia-reperfusion group(ligating the left anterior descending coronary artery as ischemia-reperfusion operation under pentobarbital anesthesia),sevoflurane group(surgery under sevoflurane anesthesia),and sevoflurane+LY294002 group(LY294002,PI3 K blocker,was injected daily one week before ischemia-reperfusion surgery and the rest was the same as in Sevoflurane group).Plasma was collected to detect the levels of myocardial injury markers,creatine kinase-MB(CK-MB)and cardiac troponin I(cTnI).Myocardial tissue was detected by western blot method to detect autophagy markers,LC3 and Beclin1,and signaling pathway proteins,phosphatase 3-kinase(PI3 K),protein kinase B(Akt),rapamycin(mTOR)total protein and phosphorylated protein levels.Results Compared with sham operation group,the levels of plasma CK-MB,cTnI,markers of cardiac autophagy LC3 and Beclin1 in the other three groups were significantly increased(P<0.05),and the ratios of phosphorylated PI3 K,Akt and mTOR to total protein significantly decreased(P<0.05).Compared with sham operation group,the total protein levels of PI3 K,Akt and mTOR in sevoflurane group were not significantly different.The levels of PI3 K and Akt significantly increased(P<0.05),and the level of mTOR protein significantly decreased(P<0.05)in ischemia-reperfusion group.The level of PI3 K in sevoflurane+LY294002 group significantly increased(P<0.05),and the protein levels of Akt and mTOR significantly decreased(P<0.05).Compared with ischemia-reperfusion group,plasma LEVELS of CK-MB and cTnI,LC3 and Beclin1 in sevoflurane group and sevoflurane+LY294002 group significantly decreased(P<0.05).Compared with the ischemia-reperfusion group,the ratio of phosphorylated PI3 K,Akt and mTOR protein to total protein levels in sevoflurane group significantly increased(P<0.05).Compared with sevoflurane group,plasma LEVELS of CK-MB,cTnI,LC3 and Beclin1 in sevoflurane+LY294002 group significantly increased(P<0.05).The levels of phosphorylated PI3 K,Akt and mTOR protein to total protein in sevoflurane+LY294002 group significantly decreased(P<0.05).Conclusions Sevoflurane inhibits the excessive autophagy induced by ischemia-reperfusion,and the possible mechanism is that it upregulates the phosphorylation level of PI3 K/Akt/mTOR pathway protein.
作者
任倪辉
郭志佳
王鑫
王成钢
张文颉
田首元
REN Nihui;GUO Zhijia;WANG Xin;WANG Chenggang;ZHANG Wenjie;TIAN Shouyuan(Department of Anesthesiology,Shanxi Medical University,Taiyuan 030001,China;Department of Anesthesiology,The First Hospital of Shanxi Medical University,Taiyuan 030001,China;Department of Anesthesia and Surgery,Shanxi Provincial Hospital of Traditional Chinese Medicine,Taiyuan 030012,China)
出处
《武警医学》
CAS
2022年第5期416-419,共4页
Medical Journal of the Chinese People's Armed Police Force
基金
山西省自然科学基金(201801D121226)
