目的 探讨七氟醚麻醉对大鼠认知功能及神经细胞凋亡的影响。方法 取96只大鼠，随机分为对照组(吸入2L·min-1氧气)、七氟醚低浓度组(吸入体积分数为1％的七氟醚+2L·min-1氧气)、七氟醚中浓度组(吸入体积分数为2％的七氟醚+2L·min-l氧气)、七氟醚高浓度组(吸入体积分数为4％的七氟醚+2L·min-1氧气)，持续吸入6h。用Morris水迷宫实验观察各组大鼠认知能力，用HE染色观察海马组织学形态，用酶联免疫吸附试验(enzyme linked immunosorbent assay，ELISA)测定海马组织中枢神经特异性蛋白(soluble protein 100β，S-100β)、神经元特异性烯醇化酶(neuron-specific enolase，NSE)、白细胞介素-1β(interleukin-1β，IL-1β)和肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)水平，流式细胞术测定神经细胞凋亡情况，蛋白印迹法(Western blotting)测定海马组织 B淋巴细胞瘤-2(B-cell lymphoma-2，Bcl-2)、Bcl-2相关 X蛋白(Bcl2-associated X protein，Bax)、磷酸化磷脂酰肌醇 3-激酶(phosphorylated phosphatidylinositol 3-kinase，p-PI3K)、磷酸化丝氨酸/苏氨酸激酶(phosphorylated serine/threonine kinase，p-Akt)和磷酸化哺乳动物雷帕霉素靶蛋白(phosphorylated mammalian target of rapamycin，p-mTOR)的水平。结果 HE染色结果表明，对照组海马组织神经细胞结构正常，排列紧密，七氟醚各浓度组大鼠海马组织神经细胞减少，排列紊乱，分布不均匀。与对照组比较，七氟醚各浓度组大鼠逃避潜伏期时间，S100-β、NSE、IL-1β、TNF-α水平，Bax蛋白水平和神经细胞凋亡率均升高，穿越平台次数，平台停留时间，Bcl-2、p-PI3K、p-Akt和p-mTOR蛋白水平均降低(P＜0。05);七氟醚各浓度组诸项指数水平变化规律相同，并呈剂量依赖性(P＜0。05)。结论 七氟醚麻醉可诱导大鼠神经细胞凋亡，使大鼠认知能力衰退，其可能与调控PI3K/Akt/mTOR信号通路有关。
Effect of sevoflurane anesthesia on neuronal apoptosis in rats based on PI3K/Akt/mTOR pathway
Objective To investigate the effect of sevoflurane anesthesia on cognitive function and nerve cells apoptosis in rats.Methods 96 rats were randomly divided into control group(inhale 2 L·min-1 oxygen),low concentration of sevaflurane group(inhale volume fraction 1％sevaflurane+2 L·min-1 oxygen),medium concentration of sevaflurane group(inhale volume fraction 2％sevaflurane+2 L·min-1 oxygen),and high concentration of sevaflurane group(inhale volume fraction 4％sevaflurane+2 L·min-1 oxygen).The inhalation lasted for 6 hours.The cognitive ability of rats was observed by Morris water maze test,and the hippocampal histological morphology was observed by HE staining.The levels of soluble soluble protein 100β(S-100β),neuron-specific enolase(NSE),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)were determined by enzyme linked immunosorbent assay(ELISA).The neuro apoptosis was determined by flow cytometry,and the levels of B-cell lymphoma-2(Bcl-2),Bcl2-associated X protein(Bax),phosphorylated phosphatidylinositol 3-kinase(p-PI3K),phosphorylated serine/threonine kinase(p-Akt)and phosphorylated mammalian target of rapamycin(p-mTOR)proteins were determined by Western blotting.Results HE staining showed the hippocampal tissue nerve cells in control group were normal and tightly arranged,the hippocampal tissue nerve cells were reduced,disordered and poorly distributed in different concentrations of sevoflurane groups.Compared with control group,the time of escape latency,S100-β,NSE,IL-1β,TNF-α,Bax protein levels and nerve cells apoptosis rates were significantly increased in different concentrations of sevoflurane groups,and the number of times through the platform,residence time on platform,Bcl-2,p-PI3K,p-Akt and p-mTOR protein levels were significantly reduced(P＜0.05).The changes of indexes in sevoflurane groups were the same,and they were dose-dependent(P＜0.05).Conclusion Sevoflurane anesthesia induces apoptosis in nerve cells and causes cognitive decline in rats,which may be related to the regulation of PI3K/Akt/mTOR signaling.
sevofluranenerve cellsapoptosisphosphoinositide 3-kinaseserine/threonine kinasemammalian target of rapamycin
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