甲基莲心碱对七氟烷麻醉后大鼠认知障碍影响的实验研究

Effect of neferine on cognitive impairment after sevoflurane anesthesia in rats

目的探讨甲基莲心碱对七氟烷麻醉后大鼠认知障碍的影响及其分子机制。方法选取2018年7月将45只SD大鼠按照随机数字表法分为对照组、模型组和甲基莲心碱组,每组15只。模型组和甲基莲心碱组大鼠七氟烷麻醉3 h后自然苏醒,对照组不做处理。甲基莲心碱组大鼠在麻醉之前给予腹腔注射甲基莲心碱5 mg/kg,对照组和模型组大鼠经腹腔注射等体积生理盐水。干预24 h,采用Morris水迷宫实验检测大鼠空间学习记忆能力;采用酶联免疫法测定3组大鼠血清氧化应激指标变化;采用原位缺口末端标记法(TUNEL)染色分析3组大鼠脑组织细胞凋亡水平;采用荧光定量聚合酶链反应(polymerase chain reaction,PCR)分析3组大鼠脑组织中半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3和Caspase-9 mRNA表达水平。应用SPSS 17.0统计软件分析,符合正态分布的计量资料以均数±标准差(±s)表示。结果甲基莲心碱组大鼠逃避潜伏期[(7.95±1.89)s]低于模型组大鼠逃避潜伏期[(12.25±2.78)s],而穿台指数(4.98±1.03)高于模型组穿台指数(3.01±0.89),差异均有统计学意义(t=3.103,P<0.05;t=1.895,P<0.05)。甲基莲心碱组大鼠血清丙二醛(MDA)水平[(8.20±1.21)μmol/L]低于模型组血清MDA水平[(13.19±1.59)μmol/L],差异有统计学意义(t=2.908,P<0.05)。甲基莲心碱组大鼠血清超氧化物歧化酶(SOD)水平[(410.43±60.12)U/ml]高于模型组血清SOD水平[(321.10±50.21)U/ml],差异有统计学意义(t=2.441,P<0.05)。甲基莲心碱组大鼠脑组织细胞凋亡比例[(5.01±1.21)%]低于模型组脑组织细胞凋亡比例[(9.10±2.19)%],差异有统计学意义(t=1.809,P<0.05)。甲基莲心碱组大鼠脑组织Caspase-3和Caspase-9 mRNA表达水平(1.39±0.15、1.52±0.19)低于模型组脑组织Caspase-3和Caspase-9 mRNA表达水平(1.98±0.23、2.42±0.25),差异有统计学意义(t=1.831,P<0.05;t=1.592,P<0.05)。结论甲基莲心碱通过下调氧化应激反应,显著缓解了七氟烷麻醉所致大鼠认知障碍。

Objective To investigate the effect of neferine on cognitive impairment after sevoflurane anesthesia in rats and its molecular mechanism.Methods A total of 45 SD rats from July 2018 to December 2019 were randomly divided into control group,model group and neferine group.Rats in the model group and neferine group recovered spontaneously after sevoflurane anesthesia for 3 h,while the control group did not receive any treatment.The rats in the neferine group were intraperitoneally injected with neferine(5 mg/kg)before anesthesia,and the rats in the control group and model group were intraperitoneally injected with the same volume of normal saline.After 24 h of intervention,Morris water maze test was used to detect the spatial learning and memory ability.Serum oxidative stress indexes and inflammatory factors were analyzed by enzyme-linked immunosorbent assay(ELISA).The apoptosis level of brain tissue in the three groups were analyzed by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining.The expression levels of cysteinyl aspartate-specific protease(Caspase-3 and Caspase-9)mRNA in the brain tissue of the three groups were analyzed by fluorescent quantitative polymerase chain reaction(PCR).Results Compared with the escape latency[(12.25±2.78)s]and perforator index(3.01±0.89)in the model group,the escape latency[(7.95±1.89)s]was significantly reduced and perforator index[(4.98±1.03)s]was significantly enhanced in the neferine group(t=3.103,1.895,P<0.05).Compared with the serum malondialdehyde(MDA)level[(13.19±1.59)μmol/L]in the model group,the serum MDA level[(8.20±1.21)μmol/L]in the neferine group was significantly decreased(t=2.908,P<0.05).Compared with the serum superoxide dismutase(SOD)level in the model group[(321.10±50.21)U/ml],the serum SOD level in the neferine group[(410.43±60.12)U/ml]was significantly increased(t=2.441,P<0.05).Compared with apoptotic rate in the model group[(9.10±2.19)%],the apoptotic rate of brain tissue in the neferine group[(5.01±1.21)%]was significantly reduced(t=1.809,P<0.05).Compared with the expression levels of Caspase-3 and Caspase-9 mRNA(1.98±0.23,2.42±0.25)in the model group,the expression levels of Caspase-3 and Caspase-9 mRNA in the neferine group(1.39±0.15,1.52±0.19)significantly decreased(t=1.831,1.592,P<0.05).Conclusion Neferine can significantly alleviate the cognitive impairment induced by sevoflurane anesthesia by down-regulating the levels of oxidative stress.