老龄雄性C57BL/6J小鼠剖腹探查术后肠道菌群及其代谢物失调与认知功能障碍的关系  

Relationship between gut microbiota and its metabolite dysregulation and postoperative cognitive dysfunction in elderly male C57BL/6J mice after laparotomy exploration

作　　者：承耀中 练新荣 李慧娴 王太行 郑晖 阎涛 Cheng Yaozhong;Lian Xinrong;Li Huixian;Wang Taihang;Zheng Hui;Yan Tao(Department of Anesthesiology,National Cancer Center,National Clinical Research Center for Cancer,Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,China;Department of Radiotherapy,National Cancer Center,National Clinical Research Center for Cancer,Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,China)

机构地区：[1]国家癌症中心,国家肿瘤临床医学研究中心,中国医学科学院北京协和医学院肿瘤医院麻醉科,北京100021 [2]国家癌症中心,国家肿瘤临床医学研究中心,中国医学科学院北京协和医学院肿瘤医院放疗科,北京100021

出　　处：《中华医学杂志》2024年第15期1316-1322,共7页National Medical Journal of China

基　　金：中国医学科学院医学与健康科技创新工程项目(2022-I2M-C&T-B-061);中国癌症基金会北京希望马拉松专项(LC2021A18,LC2022B06)

摘　　要：目的探讨老龄雄性C57BL/6J小鼠剖腹探查术后认知功能障碍与肠道菌群及其代谢物失调改变的关系。方法采用随机数字表法将48只老龄(16~17月龄)无特定病原体(SPF)雄性C57BL/6J小鼠分为对照组(n=24)和手术组(n=24)。手术组采用1.4%异氟醚诱导麻醉15 min后,继续用1.4%异氟醚+100%氧气维持麻醉行剖腹探查术,手术时间持续10 min,术后继续麻醉2 h;对照组不予手术及麻醉,置于含100%氧气的透明箱中2 h。术后48 h同时采集两组小鼠的粪便和外周血。采用16S rDNA基因测序检测肠道菌群的丰度、多样性和差异性,采用液相色谱-串联质谱(LC/MS)代谢组技术检测粪便代谢谱的改变,并分析差异代谢物功能,采用ELISA检测血清白细胞介素(IL)-6、IL-1β和肿瘤坏死因子-α(TNF-α)含量。在术后3 d行Morris水迷宫实验,检测小鼠认知功能。结果对照组与手术组小鼠术后逃避潜伏期分别为(22.0±4.9)、(35.0±5.1)s,目标象限停留时间分别为(26.0±3.7)、(16.0±2.9)s;与对照组比较,手术组小鼠术后逃避潜伏期延长(P=0.035),目标象限停留时间减少(P=0.006)。两组小鼠肠道菌群差异变化具有可比性,手术组大肠埃希菌、志贺菌和梭杆菌的菌群表达水平上调,瘤胃菌和丁酸菌的菌群表达水平下调。LC/MS获得对照组和手术组小鼠粪便代谢谱,在正、负离子模式分别筛选出14种和21种差异代谢物,正离子模式差异代谢物为谷氨酸、2-吲哚甲酸、犬尿酸、甘油醛等,负离子模式差异代谢物为甲硫氨酸、天冬氨酸、L-苏氨酸、酪氨酰-苏氨酸、5-羟基吲哚-3-乙酸等。鉴定出的差异代谢物的路径主要涉及氨基酸、脂肪酸和色氨酸代谢以及核苷酸合成。两组小鼠血清IL-1β、IL-6和TNF-α表达水平差异均无统计学意义(均P>0.05)。结论肠道菌群及其代谢物失调与老龄雄性C57BL/6J小鼠术后认知功能障碍的发生相关。麻醉手术不仅改变了小鼠肠道微�Objective To explore the relationship between gut microbiota and its metabolite dysregulation and postoperative cognitive dysfunction in elderly male C57BL/6J mice after laparotomy exploration.Methods A total of 48 specific pathogen-free(SPF)male C57BL/6J mice,aged 16-17 months,were divided into two groups by random number table method:control group(n=24)and operation group(n=24).Mice in the operation group were induced with 1.4%isoflurane for 15 minutes,followed by a 10 minutes exploratory laparotomy anesthetized with 1.4%isoflurane and 100%oxygen,and anesthesia continued for 2 hours after surgery.Mice in control group were put in 100%oxygen for 2 hours.Feces and venous blood samples of both groups were collected 48 hours after surgery.Changes in the abundance and diversity of intestinal bacteria in the feces were detected by 16S rDNA gene sequencing.Functional changes of fecal metabolic profiles were detected by liquid chromatography tandem mass spectrometry(LC/MS)metabolomics and differential metabolite functions were analyzed.The serum level of interleukin(IL)-6,IL-1βand tumor necrosis factor-α(TNF-α)were detected by Enzyme-linked immunosorbent assay(ELISA).The cognitive function of the mice was detected by Morris water maze test 3 days after operation.Results The postoperative escape latency of mice in control group and operation group was(22.0±4.9)and(35.0±5.1)s,and the target quadrant residence time was(26.0±3.7)and(16.0±2.9)s,respectively.Compared with the control group,the postoperative escape latency of mice in the operation group was prolonged(P=0.035),and the residence time in the target quadrant was reduced(P=0.006).The difference of intestinal flora between the two groups was comparable.The expression levels of Escherichia coli,shigella and clostridium in the operation group were up-regulated,while the expression levels of rumen bacteria and butyricobacteria were down-regulated.Fecal metabolic profiles of mice in control group and operation group were obtained by LC/MS,and 14 and 21 different

关 键 词：术后认知功能障碍 肠道菌群失调 微生物-肠-脑轴 代谢组 16S rDNA测序 

分 类 号：R614[医药卫生—麻醉学]