基于非靶向代谢组学分析肠道代谢物在术后认知功能中的作用  

The role of intestinal metabolites in postoperative cognitive function was analyzed based on non-targeted metabolomics

作　　者：张世花 伍晴 韩俊刚 徐龙生 贾晓玉 周清河[3] Zhang Shihua;Wu Qing;Han Jungang;Xu Longsheng;Jia Xiaoyu;Zhou Qinghe(College of Life Science and Medicine,Zhejiang Sci-Tech University,Hangzhou 310018,China;Central Laboratory,the Affiliated Hospital of Jiaxing University,Jiaxing 314000,China;Department of Anesthesia and Surgery Center,the Affiliated Hospital of Jiaxing University,Jiaxing 314000,China)

机构地区：[1]浙江理工大学生命科学与医药学院,杭州310018 [2]嘉兴学院附属医院中心实验室,嘉兴314000 [3]嘉兴学院附属医院麻醉手术中心,嘉兴314000

出　　处：《中华实验外科杂志》2023年第11期2257-2261,共5页Chinese Journal of Experimental Surgery

基　　金：浙江省基础公益研究计划(LGF21H090017);浙江省中医药多学科交叉创新团队(2022-19);嘉兴市医学重点学科(2023-ZC-001)。

摘　　要：目的:探究肠道代谢物紊乱在术后认知功能障碍的发生中的作用及其机制。方法:将18月龄的C57BL/6雄性小鼠,采用随机数字表法将动物分为空白对照组、麻醉/手术(1.5~2.0%异氟醚麻醉2 h持续3 d+腹腔探查术组)、油酸酰胺组[(5 mg/(kg·d)]。模拟临床过程建立麻醉/手术诱导的老年术后认知功能障碍模型。使用Morris水迷宫实验和旷场实验检测小鼠麻醉/手术前和麻醉/手术后3 d认知水平变化。对发生认知功能障碍的麻醉/手术组小鼠和空白对照组小鼠的结肠远端内容物进行非靶向代谢组学分析。将获得的代谢组学原始数据进行生物信息学分析比较麻醉/手术组与对照组之间的差异代谢物。选取内源性差异代谢物之一油酸酰胺进行补充。两组间比较采用非配对t检验,多组间比较采用单因素方差分析(ANOVA)。结果:Morris水迷宫实验结果显示麻醉/手术组小鼠逃避潜伏期较对照组明显延长[(24.3±3.0)s比(8.3±1.3)s,P<0.05],靶向象限探索时间缩短[(24.1±4.6)s比(44.0±4.2)s,P<0.05],两组小鼠平均游泳速度差异无统计学意义[(205.3±28.9)mm/s比(214.6±8.9)mm/s,P>0.05]。在旷场实验中麻醉/手术组小鼠相比于对照组总活动路程减少[(3636.6±539.6)mm比(12398.4±2812.5)mm,P<0.05],中央探索时间缩短[(11.7±2.7)s比(63.1±9.3)s,P<0.05],平均速度减慢[(10.6±2.0)mm/s比(21.7±4.8)mm/s,P<0.05],静止时间延长[(464.8±41.3)mm/s比(388.9±51.7)mm/s,P<0.05]。非靶向代谢组学分析结果显示麻醉/手术组与空白对照组之间存在35种差异代谢物(VIP>1,P<0.05)。老年小鼠接受油酸酰胺补充后学习记忆能力明显改善,在Morris水迷宫实验中油酸酰胺组小鼠逃避潜伏期相对于麻醉/手术组明显缩短[(14.9±2.8)s比(22.2±5.2)s,P<0.05],对靶向象限的偏好性增强[(36.2±2.3)s比(20.0±3.8)s,P<0.05]。结论:老年术后认知功能障碍的发生与肠道内源性代谢物紊乱有关。Objective To investigate the association between post-operative cognitive dysfunction and changes in gut metabolites.Methods C57BL/6 male mice of 18 months of age were numbered one by one,and the animals were divided into blank control group,anesthesia/surgery group(1.5-2.0%isoflurane anesthesia for 2 h lasting for 3 d+laparotomy group),and oleic acid amide group(5 mg/kg/day)by using random number table method.A model of anesthesia/surgery-induced postoperative cognitive dysfunction in the elderly was established by simulating the clinical process.Changes in cognitive levels in mice before and 3d after anesthesia/surgery were detected using the Morris water maze experiment and the open field experiment.Non-targeted metabolomics was performed on the distal colon contents of mice in the anesthesia/surgery group that developed cognitive dysfunction and mice in the blank control group.The obtained metabolomics raw data were subjected to bioinformatics analysis to compare the differential metabolites between the anesthetized/surgical group and the control group.One of the endogenous differential metabolites,oleamide,was selected for supplementation.Unpaired t-tests were used for comparisons between two groups,and one-way ANOVA was used for comparisons between multiple groups.Results The results of Morris water maze experiments showed that the escape latency of mice in the anesthetized/surgical group was significantly longer than that in the control group[(24.3±3.0)s vs.(8.3±1.3)s,P<0.05],and the time of exploration of the target quadrant was shorter[(24.1±4.6)s vs.(44.0±4.2)s,P<0.05],there was no difference in the mean swimming speed of the mice in the two groups[(205.3±28.9)mm/s vs.(214.6±8.9)mm/s,P>0.05].In the open field experiment mice in the anesthetized/surgical group had reduced total active distance traveled compared to the control group[(3636.6±539.6)mm vs.(12398.4±2812.5)mm,P<0.05],shorter central exploration time[(11.7±2.7)s vs.(63.1±9.3)s,P<0.05],slower mean velocity[(10.6±2.0)mm/s vs.(21.7±4.8

关 键 词：麻醉/手术 术后认知功能障碍 代谢组学 油酸酰胺 

分 类 号：R614[医药卫生—麻醉学]