七氟醚通过海马PICK1/ERK1/2/GSK3β信号抑制记忆巩固的研究
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摘要
目的研究七氟醚对记忆巩固的影响,并探讨PICK1/ERK1/2/GSK3β信号通路在其中的作用。方法采用C57BL/6J PICK1基因敲除小鼠进行七氟醚处理(2%,3 h),提取动物海马组织蛋白,采用Western blot检测PICK1、ERK1/2、p-ERK1/2、GSK3β及p-GSK3β蛋白表达变化。采用原代海马神经细胞,加入ERK1/2磷酸化激动剂Honokiol和/或PICK1抑制剂FSC231,再进行七氟醚处理(2%,3 h),检测p-GSK3b蛋白表达变化。通过旷场试验检测动物运动能力变化,抑制性逃避试验检测动物记忆巩固的变化。结果在体实验中,单独进行七氟醚处理或敲除PICK1基因均可明显降低PICK1、p-ERK1/2及p-GSK3β的蛋白表达水平。与七氟醚组相比,PICK1基因敲除+七氟醚处理组,p-ERK1/2及p-GSK3β的蛋白表达水平进一步降低。离体实验中,七氟醚可降低p-GSK3β蛋白表达水平,加入Honokiol可使p-GSK3β蛋白表达增加,而再加入FSC231则可逆转由Honokiol引起的p-GSK3β蛋白变化。行为学结果显示,七氟醚处理或PICK1基因敲除动物运动能力无明显变化,而七氟醚可抑制记忆巩固;PICK1基因敲除小鼠进行七氟醚处理,则可加重抑制记忆巩固。结论七氟醚可抑制记忆巩固过程,其机制可能与PICK1/ERK1/2/GSK3β信号通路受抑制相关。
OBJECTIVE To study the effects of sevoflurane on memory consolidation, and the role of PICK1/ERK1/2/GSK3β signaling pathway. METHODS C57 BL/6 J PICK1 knock-out mice were exposed to 2% sevoflurane for 3 h. The hippocampal tissues were collected for detecting the protein expressions of PICK1, ERK1/2, P-ERK1/2, GSK3β and P-GSK3β by Western blot. Primary hippocampal neurons were treated with phospho-ERK1/2 activator Honokiol or/and PICK1 inhibitor FSC231 before sevoflurane(2%, 3 h) exposure. Neurons were harvested for detection of p-GSK3β protein level. The open-field test and inhibitory avoidance task were enrolled for evaluation of motor activity and memory consolidation respectively. RESULTS The protein levels of PICK1, p-ERK1/2 and p-GSK3β were depressed in sevoflurane or PICK1 knock-out group. Compared with sevoflurane group, the expressions of p-ERK1/2 and p-GSK3β were further decreased in sevoflurane plus PICK1 knock-out group. The expression of p-GSK3β was inhibited by sevoflurane exposure, which could be improved by application of Honokiol. Moreover, the improvement of p-GSK3β by Honokiol could be reversed by further addition of FSC231. Finally, analysis of behaviour study showed that sevoflurane treatment or PICK1 knock-out did not alter the motor activity. However, sevoflurane administration inhibited memory consolidation. For the knock-out mice, the memory consolidation was further suppressed by sevoflurane exposure. CONCLUSION Sevoflurane can inhibit the memory consolidation process, and the mechanism may be related to inhibition of the PICK1/ERK1/2/GSK3β signaling pathway.
OBJECTIVE To study the effects of sevoflurane on memory consolidation, and the role of PICK1/ERK1/2/GSK3β signaling pathway. METHODS C57 BL/6 J PICK1 knock-out mice were exposed to 2% sevoflurane for 3 h. The hippocampal tissues were collected for detecting the protein expressions of PICK1, ERK1/2, P-ERK1/2, GSK3β and P-GSK3β by Western blot. Primary hippocampal neurons were treated with phospho-ERK1/2 activator Honokiol or/and PICK1 inhibitor FSC231 before sevoflurane(2%, 3 h) exposure. Neurons were harvested for detection of p-GSK3β protein level. The open-field test and inhibitory avoidance task were enrolled for evaluation of motor activity and memory consolidation respectively. RESULTS The protein levels of PICK1, p-ERK1/2 and p-GSK3β were depressed in sevoflurane or PICK1 knock-out group. Compared with sevoflurane group, the expressions of p-ERK1/2 and p-GSK3β were further decreased in sevoflurane plus PICK1 knock-out group. The expression of p-GSK3β was inhibited by sevoflurane exposure, which could be improved by application of Honokiol. Moreover, the improvement of p-GSK3β by Honokiol could be reversed by further addition of FSC231. Finally, analysis of behaviour study showed that sevoflurane treatment or PICK1 knock-out did not alter the motor activity. However, sevoflurane administration inhibited memory consolidation. For the knock-out mice, the memory consolidation was further suppressed by sevoflurane exposure. CONCLUSION Sevoflurane can inhibit the memory consolidation process, and the mechanism may be related to inhibition of the PICK1/ERK1/2/GSK3β signaling pathway.
引文
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[10]ALKIRE M T,GRUVER R,MILLER J,et al.Neuroimaging analysis of an anesthetic gas that blocks human emotional memory[J].Proc Natl Acad Sci USA,2008,105(5):1722-1727.
[11]XU Z,DONG Y,WANG H,et al.Age-dependent postoperative cognitive impairment and Alzheimer-related neuropathology in mice[J].Sci Rep,2014(4):3766.
[2]GONZALEZ DE VALDIVIA E,BROSELID S,KAHN R,et al.G protein-coupled estrogen receptor 1(GPER1)/GPR30increases ERK1/2 activity through PDZ motif-dependent andindependent mechanisms[J].J Biol Chem,2017,292(24):9932-9943.
[3]YANG S Y,CHEN W H,JIN H Z,et al.Dexmedetomidine prevents sevoflurane neurotoxicity via ERK1/2 MAPKsignaling in the developing brain[J].Chin J Mod Appl Pharm(中国现代应用药学),2014,31(5):523-528.
[4]NEEDHAM M J,WEBB C E,BRYDEN D C.Postoperative cognitive dysfunction and dementia:what we need to know and do[J].Br J Anaesth,2017,119(Suppl 1):i115-i125.
[5]VUTSKITS L,XIE Z C.Lasting impact of general anaesthesia on the brain:mechanisms and relevance[J].Nat Rev Neurosci,2016,17(11):705-717.
[6]XING B,HAN G N,WANG M J,et al.Juvenile treatment with m GluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β[J].Neuropharmacology,2018(137):359-371.
[7]NAGARAJAN D,MELO T,DENG Z Y,et al.ERK/GSK3β/Snail signaling mediates radiation-induced alveolar epithelial-to-mesenchymal transition[J].Free Radic Biol Med,2012,52(6):983-992.
[8]LIN E Y S,SILVIAN L F,MARCOTTE D J,et al.Potent PDZ-domain PICK1 inhibitors that modulate amyloid beta-mediated synaptic dysfunction[J].Sci Rep,2018(8):13438.
[9]ALKIRE M T,NATHAN S V,MCREYNOLDS J R.Memory enhancing effect of low-dose sevoflurane does not occur in basolateral amygdala-lesioned rats[J].Anesthesiology,2005,103(6):1167-1173.
[10]ALKIRE M T,GRUVER R,MILLER J,et al.Neuroimaging analysis of an anesthetic gas that blocks human emotional memory[J].Proc Natl Acad Sci USA,2008,105(5):1722-1727.
[11]XU Z,DONG Y,WANG H,et al.Age-dependent postoperative cognitive impairment and Alzheimer-related neuropathology in mice[J].Sci Rep,2014(4):3766.