金刚烷胺减轻海马炎症反应和细胞凋亡在小鼠术后认知功能障碍中的治疗作用

Role of amantadine alleviating hippocampal inflammationand apoptosis in postoperative cognitive dysfunction in mice

目的探讨非竞争性N-甲基-M-天冬氨酸受体拮抗剂金刚烷胺对术后认知功能障碍(POCD)的治疗作用及可能机制。方法雄性C57BL/6小鼠54只,16月龄,体重35~40 g,随机分为三组,每组18只:对照组(CV组)、七氟醚麻醉+剖腹探查+溶剂组(PV组)、七氟醚麻醉+剖腹探查+金刚烷胺组(PA组)。采用七氟醚麻醉+剖腹探查术建立POCD动物模型。PA组于麻醉前腹腔注射金刚烷胺25 mg/kg,之后每日注射一次直到术后第2天;另外两组注射等容量溶剂。术后第6天行旷场实验,第7天行条件性恐惧实验训练,第8天行条件性恐惧测试。术后第1天采用Western blot检测海马组织白细胞介素-1β(IL-1β)和活化的半胱氨酸天冬氨酸蛋白酶-3(cleaved caspase-3)含量,免疫荧光染色结合图像分析技术检测神经核抗原(NeuN)阳性细胞数量。结果三组总探索路程和中央格停留时间差异无统计学意义。与CV组比较,PV组僵直时间明显缩短,IL-1β、cleaved caspase-3含量明显升高,NeuN阳性细胞数量明显减少(P<0.05)。与PV组比较,PA组僵直时间明显延长,IL-1β、cleaved caspase-3含量明显降低,NeuN阳性细胞数量明显增多(P<0.05)。结论金刚烷胺可能通过减少海马炎症反应和细胞凋亡来改善POCD。

Objective To investigate the therapeutic effect and possible mechanisms of non-competitive N-methyl-M-aspartate receptor antagonist amantadine on postoperative cognitive dysfunction(POCD).Methods Fifty-four male C57 BL/6 mice,aged 16 months,weighing 30-40 g,were randomly divided into three groups(n=18):control group(group CV),sevoflurane anesthesia+surgery+vehicle group(group PV),sevoflurane anesthesia+surgery+amantadine group(group PA).POCD animal model was established by sevoflurane anesthesia and exploratory laparotomy.In group PA,mice were intraperitoneally injected with amantadine 25 mg/kg before anesthesia,and then injected once a day till the second day after surgery.The mice in the other two groups were injected with an equal volume of vehicle.Open field was performed on the 6 th day after surgery,the training of fear conditioning was performed on the 7 th day,and the contextual fear conditioning test was performed on the 8 th day.The hippocampus tissues of some mice were collected on the first day after surgery.The expressions of interleukin-1β(IL-1β)and cleaved caspase-3 were detected by Western blot.The number of NeuN positive cells was detected by using immunofluorescence staining combined image analysis technique.Results No significant difference was observed in the total travel distance and the time spent in the center of the area between the three groups.Compared with group CV,the freezing time to context was significantly decreased while the expressions of IL-1βand cleaved caspase-3 were significantly increased and the number of NeuN positive cells decreased in the hippocampus in group PV(P<0.05).Compared with group PV,the freezing time to context was significantly increased while the expressions of IL-1βand cleaved caspase-3 were significantly decreased and the number of NeuN positive cells increased in the hippocampus in group PV(P<0.05).Conclusion AMA may improve the cognitive impairment by alleviating hippocampal inflammation and apoptosis.