CGRP通过调控PI3K/Akt/mTOR信号通路对七氟醚诱导新生大鼠神经毒性的影响  

Effect of CGRP on sevoflurane-induced neurotoxicity in neonatal rats by regulating PI3K/Akt/mTOR signaling pathway

作　　者：李兔平 韩毅[1] 孙韬[1] 雷易 刘臻 贾丽玲 张林忠[1] Li Tuping;Han Yi;Sun Tao;Lei Yi;Liu Zhen;Jia Liling;Zhang Linzhong(Department of Anesthesiology,Shanxi Medical University Second Hospital,Shanxi,Taiyuan 030001,China)

机构地区：[1]山西医科大学第二医院麻醉科,太原030001

出　　处：《疑难病杂志》2025年第4期485-491,共7页Chinese Journal of Difficult and Complicated Cases

基　　金：山西省基础研究计划青年科学研究项目(202203021222392)。

摘　　要：目的探讨降钙素基因相关肽(CGRP)对七氟醚诱导的新生大鼠神经毒性及磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的影响。方法于2023年1月—2024年6月在山西医科大学第二医院麻醉研究室进行实验。构建七氟醚诱导的新生大鼠模型,将造模大鼠随机分为模型组(Model组)、CGRP组、CGRP+LY294002组,每组12只,另取12只正常大鼠作对照组(Control组);Morris水迷宫实验检测各组大鼠认知功能;酶联免疫吸附试验(ELISA)检测氧化应激水平;苏木素—伊红染色法(HE)检测脑组织病理损伤情况;缺口末端标记法(TUNEL)检测神经元凋亡情况;免疫组化检测自噬相关蛋白表达;免疫印记法(Western blot)检测PI3K/Akt/mTOR信号通路及凋亡相关蛋白表达。结果与Control组比较,Model组脑组织结构遭到破坏,神经元排列紊乱,数量减少,核固缩深染;大鼠逃逸潜伏期延长、目标象限停留时间缩短,丙二醛(MDA)水平及神经元凋亡率、B淋巴细胞瘤-2(Bcl-2)相关X蛋白(Bax)、重组人自噬效应蛋白(Beclin1)表达升高,超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-px)水平及Bcl-2、p62、磷酸化(p)-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR表达降低(P<0.01或P<0.05)。与Model组比较,CGRP组脑组织结构相对正常,神经元排列相对整齐,少量神经元凋亡,核固缩现象明显减轻;大鼠逃逸潜伏期缩短,目标象限停留时间延长,MDA水平及神经元凋亡率、Bax、Beclin1表达降低,SOD、GSH-px水平及Bcl-2、p62、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR表达升高(P<0.01或P<0.05)。与CGRP组比较,CGRP+LY294002组脑组织破坏严重,神经元病理损伤加重;逃逸潜伏期延长,目标象限停留时间缩短,MDA水平及神经元凋亡率、Bax、Beclin1表达升高,SOD、GSH-px水平及Bcl-2、p62、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR表达降低(P<0.01或P<0.05)。结论CGRP可减轻七氟醚诱导新生大鼠神经毒性,其作用机制�Objective To investigate the effects of calcitonin gene-related peptide(CGRP)on sevoflurane-induced neurotoxicity and phosphatidylinositol 3-kinase(PI3k)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway in neonatal rats.Methods Conducted the experiment in the Anesthesia Research Laboratory of the Second Hospital of Shanxi Medical University from January 2023 to June 2024.The neonatal rat model induced by sevoflurane was constructed,the model rats were randomly divided into model group(Model group),CGRP group and CGRP+LY294002 group,with 12 rats in each group,another 12 normal rats were taken as control group(Control group).The cognitive function of rats in each group were detected by Morris water maze.The level of oxidative stress was detected by enzyme-linked immunosorbent assay(ELISA).The pathological damage of brain tissue was detected by hematoxylin-eosin(HE)staining.Neuronal apoptosis was detected by terminal deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining.Through the use of immunohistochemistry,the expression of proteins associated to autophagy was found.The PI3k/Akt/mTOR signaling pathway and apoptosis-related proteins were detected by Western blot.Results The brain tissue structure of the Model group was destroyed compared with Control group,the arrangement of neurons was disordered,the number of neurons was reduced,the nuclear pyknosis was deeply stained,the escape latency of rats was prolonged,the residence time in the target quadrant was shortened,the malondialdehyde(MDA)level and neuronal apoptosis rate,the expression of B-cell lymphoma-2(Bcl-2)-related X protein(Bax)and recombinant human autophagy effector protein(Beclin1)were increased,the levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-px)and the expression of Bcl-2,p62,Phosphorylated(p)-Akt/PI3k,p-Akt/Akt,p-mTOR/mTOR were decreased(P<0.05).The brain tissue structure of CGRP group was relatively normal compared with Model group,the neurons were arranged relatively neatly,a small amount

关 键 词：七氟醚 神经毒性 降钙素基因相关肽 磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路 大鼠 

分 类 号：R741.05[医药卫生—神经病学与精神病学]