目的 通过粪菌移植探究肠道菌群是否参与手术创伤及麻醉引起的围术期神经认知紊乱。 方法 将36只C57BL/6J小鼠随机分为对照组（Control组）、手术组（Surgery组）和粪菌移植组（fecal microbiota transplantation, FMT组）。小鼠在全麻下行肝部分切除术制备围术期神经认知紊乱模型，采集手术组小鼠术后粪便制成粪菌滤液移植至FMT组小鼠。采用蛋白免疫印迹法检测小鼠大脑海马区IL-1β、IL-6、脑源性神经营养因子（brain-derived neurotrophic factor, BDNF）的表达水平，免疫组织化学法检测小胶质细胞激活标志物——离子钙接头蛋白抗原（Ionized calcium bindingadaptor molecule-1, Iba-1）的表达高低，通过Morris水迷宫（Morris water maze, MWM）评估认知功能。 结果 与对照组相比，手术组小鼠在术后探索过程中逃逸潜伏期延长，穿越平台次数减少（P＜0.001），在目标象限内的游泳时间缩短（P＜0.001），IL-1β、IL-6在术后第3天的表达水平与对照组相比显著升高（P＜0.001, P＜0.001），小胶质细胞激活增加（P＜0.001），BDNF则有所下降（P＜0.05）；与对照组相比，粪菌移植组小鼠出现空间学习和记忆能力损伤（P＜0.001），IL-1β、IL-6、Iba-1在移植后第3天的表达水平也显著高于对照组（P＜0.001, P＜0.001, P＜0.001），并部分抑制了BDNF的表达（P＜0.001）。 结论 肠道菌群参与了手术创伤及麻醉引起的围术期神经认知紊乱，其机制可能与海马区神经炎症的发生、小胶质细胞的激活增加有关。
Objective To investigate whether gut microbiota is involved in surgery and anesthesia-induced perioperative neurocognitive disorders through fecal microbiota transplantation. Methods A total of 36 C57BL/6J mice were randomly divided into control (C) group, surgery (S) group and fecal microbiota transplantation (FMT) group. Mice were subjected to partial hepatectomy under general anesthesia to prepare a model of perioperative neurocognitive disorders. Fecal microbiota filtrate was collected from postoperative feces of mice in surgery group and transplanted into FMT mice. Western Blot was used to detect the levels of IL-1β, IL-6 and brain-derived neurotrophic factor (BDNF). The expression of microglia markers--Ionized calcium bindingadaptor molecule-1 (Iba-1) was measured by immunohistochemical staining. Behavioral performance was evaluated by Morris water maze (MWM). Results The escape latency of surgical mice was prolonged than the controls. The times of crossing platform were decreased (P＜0.001) and the duration of swimming in the target quadrant (P＜0.001) was shortened in surgical mice. The expression of IL-1β, IL-6 and Iba-1 were significantly increased than those in control group on postoperative day 3 (P＜0.001, P＜0.001, P＜0.001). The level of BDNF was decreased following surgery procedure (P＜0.05). The mice in FMT group showed behavioral deficits compared with the controls (P＜0.001). The levels of IL-1β, IL-6 and Iba-1 were significantly increased than the controls on post-transplantation day 3 (P＜0.001, P＜0.001, P＜0.001), and the expression of BDNF was partially inhibited (P＜0.001). Conclusions Gut microbiota involved in surgery and anesthesia-induced perioperative neurocognitive disorders, which may be related to hippocampal neuroinflammation and microglial overactivation.
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