孕期罗哌卡因麻醉通过NGF/ERK/CREB途径影响SD模型子代情绪及学习记忆能力

Ropivacaine anesthesia during pregnancy affects the emotion and learning and memory ability of SD model offspring through NGF/ERK/CREB pathway

目的探讨孕期罗哌卡因麻醉对子代情绪及学习记忆能力的影响及其潜在机制。方法本研究建立了怀孕大鼠模型,以研究在妊娠中期罗哌卡因麻醉后对后代的神经认知影响。在出生后第几天(P25~30)对其行为进行测试(Morris水迷宫和条件性恐惧实验)。此外,通过Nissl染色检测后代的海马形态(P30),并通过Golgi染色测定海马树突棘密度。通过蛋白质印迹法测定海马神经生长因子(NGF)、细胞外信号调节激酶(ERK)、磷酸化ERK(p-ERK),环磷酸腺苷反应元件结合(CREB)、p-CREB、突触素(SYP)、突触蛋白酶(SYN)和突触后密度-95(PSD95)的蛋白水平。此外,SCH772984(ERK抑制剂)用于评估PC12细胞中ERK途径的作用和潜在机制。结果结果显示罗哌卡因麻醉会导致长期神经认知功能障碍,降低树突旋转的密度,导致神经元损失,并下调海马中NGF、ERK、p-ERK、促分裂原和应激激活蛋白激酶(MSK)、CREB、p-CREB、SYP、SYN和PSD95的表达。结论在胎儿大脑发育期间,罗哌卡因诱导的母体麻醉会导致后代长期的神经认知功能障碍,这可能是通过抑制海马中的NGF/ERK/CREB通路而发生的。

Objective To investigate the effect of ropivacaine anesthesia during pregnancy on offspring’s emotion and learning and memory ability and its potential mechanism.Methods A pregnant rat model was established to investigate the neurocognitive effects of ropivacaine anesthesia on offspring during the second trimester.The behavior was tested on the first few days after birth(P25-30)(Morris water maze and conditioned fear test).In addition,the hippocampal morphology(P30)of the offspring was detected by Nissl staining,and the density of hippocampal dendritic spines was determined by Golgi staining.Hippocampal nerve growth factor(NGF),extracellular signal-regulated kinase(ERK)and phosphorylated-ERK(p-ERK)were determined by Western blotting.The protein levels of yclic adenosine monophosphate response element-binding(CREB),p-CREB,synaptophysin(SYP),synapsin(SYN)and postsynaptic density-95(PSD95).In addition,SCH772984(an ERK inhibitor)was used to evaluate the role and potential mechanism of the ERK pathway in PC12 cells.Results The results showed that ropivacaine anesthesia could lead to long-term neurocognitive dysfunction,reduce the density of dendritic rotation,lead to neuronal loss,and down-regulate the expression of NGF,ERK,p-ERK,mitogen and stress-activated protein kinase(MSK),CREB,p-CREB,SYP,SYN and PSD95 in hippocampus.Conclusion Ropivacaine-induced maternal anesthesia during fetal brain development can lead to long-term neurocognitive dysfunction in offspring,which may occur by inhibiting the NGF/ERK/CREB pathway in the hippocampus.