肠道菌群在手术及麻醉引起的围手术期神经认知紊乱中的作用  

The role of gut microbiota in surgery and anesthesia-induced perioperative neurocognitive disorders in C57BL/6J mice

作　　者：徐晓涵 周永健 李哲[1] 曹学照[1] Xu Xiaohan;Zhou Yongjian;Li Zhe;Cao Xuezhao(Department of Anesthesiology,the First Hospital of China Medical University,Shenyang 110001,China)

机构地区：[1]中国医科大学附属第一医院麻醉科,沈阳110001

出　　处：《国际麻醉学与复苏杂志》2020年第10期933-940,共8页International Journal of Anesthesiology and Resuscitation

基　　金：辽宁省自然科学基金(20180550022)。

摘　　要：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手术期神经认知紊乱(perioperative neuro-cognitive disorders,PND)。方法将36只C57BL/6J小鼠采用随机数字表法分为对照组(C组)、手术组(S组)和粪菌移植组(F组),每组12只。S组小鼠在全身麻醉下行肝部分切除术制备PND模型,采集小鼠术后粪便制成粪菌滤液移植至F组小鼠体内。采用Western blot法检测小鼠术后/粪菌移植后第3、7天大脑海马区IL-1β、IL-6、脑源性神经营养因子(brain-derived neuro-trophic 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-binding adapter molecule-1,Iba-1)的表达情况,通过Morris水迷宫(Morris water maze,MWM)评估术后/粪菌移植后第3、7天的认知功能。结果与C组比较:S组术后第3天、F组移植后第3天在目标象限内停留时间缩短(P均<0.01),穿越平台次数减少(P均<0.01);S组在术后第7天、F组在移植后第7天在目标象限内停留时间延长(P均>0.05),穿越平台次数增多(P均>0.05)。S组IL-1β、IL-6水平在术后第3天(P均<0.01)和术后第7天(分别为P<0.01、P<0.05)均较C组明显升高;F组IL-1β和IL-6水平在移植后第3天也较C组明显升高(P均<0.01),移植后第7天仍保持上调(P均<0.05)。与C组比较:S组小鼠大脑海马区小胶质细胞Iba-1水平在术后第3天明显升高(P<0.01),术后第7天仍保持上调(P<0.05);F组在移植后第3天Iba-1水平也明显提高(P<0.01),移植后第7天相差不大(P>0.05)。与C组比较:S组在术后第3天、F组在移植后第3天BDNF的表达明显降低(分别为P<0.05、P<0.01);在术后第7天S组的BDNF水平较术后第3天升高(P>0.05),移植后第7天F组仍保持在较低水平(P<0.05)。结论肠道菌群参与了手术创伤及麻醉引起的PND,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(PND)through fecal microbiota transplantation.Methods A total of 36 C57BL/6J mice were randomly divided into the control(C)group,the surgery(S)group and the fecal microbiota transplantation(FMT)group.The mice were subjected to partial hepatectomy under general anesthesia to prepare a model of PND.Fecal microbiota filtrates were collected from postoperative feces of mice in surgery group and were transplanted into FMT mice.On days 3 and 7 after postoperative or post-transplantation,the mice were sacrificed and tissues were collected.Then western blot was used to detect the protein expression levels of interleukin(IL)-1β,IL-6 and brain-derived neurotrophic factor(BDNF).The expression of microglia markers--ionized calcium binding adaptor molecule-1(Iba-1)was measured by immunohistochemical staining.Behavioral performance was evaluated by Morris water maze(MWM)on postoperative or post-transplantation days 3 and 7.Results Compared with group C,the number of platform crossings was decreased(P<0.01)and the duration of swimming in the target quadrant was shortened(P<0.01)in group S on postoperative day 3 and group F on post-transplantation day 3.On postoperative and post-transplantation day 7,the number of platform crossings was increased(P>0.05)and the duration of swimming in the target quadrant was prolonged(P>0.05)in group S and group F.The protein expression levels of IL-1βand IL-6 in surgical mice were significantly higher than the expression levels of control mice on postoperative days 3(P<0.01)and 7(P<0.01,P<0.05).The levels of IL-1βand IL-6 in group F were also significantly higher on post-transplantation day 3(P<0.01)than those in group C,and remained upregulated on post-transplantation day 7(P<0.05).Compared with group C,the level of Iba-1 in hippocampal area of mice in group S was significantly increased on postoperative day 3(P<0.01),and was still higher than the level of grou

关 键 词：围手术期神经认知紊乱 神经炎症 小胶质细胞 肠道菌群 

分 类 号：R614[医药卫生—麻醉学]