N-甲基-D-天冬氨酸受体在大鼠术后疲劳综合征的中枢发生机制研究  

Relationship between NMDA receptor and postoperative fatigue syndrome and its associated central mechanism

作　　者：陈伟哲[1] 刘舒[1] 陈帆风 周崇俊[1] 庄成乐[1] 邵时杰 俞健[1] 黄冬冬[1] 陈必成[2] 余震[3] 

机构地区：[1]温州医科大学附属第一医院胃肠外科,325000 [2]温州医科大学附属第一医院外科实验室,325000 [3]上海同济大学附属第十人民医院胃肠外科

出　　处：《中华胃肠外科杂志》2015年第4期376-381,共6页Chinese Journal of Gastrointestinal Surgery

基　　金：国家自然科学基金（81171857）;浙江省自然科学基金（LZl2H07001）;浙江省卫生部共建项目（201231118）;浙江省卫生厅医学支撑学科一营养学（11-ZC24）

摘　　要：目的：探讨N-甲基-D-天冬氨酸（NMDA）受体在大鼠术后疲劳综合征（POFS）的中枢发生机制。方法将96只SD大鼠按体质量大小编号，完全随机分为对照组（开腹后仅翻动肠袢、腹腔注射1 ml／kg 0．9％氯化钠溶液1次）、POFS模型组（70％中段小肠切除、腹腔注射1 ml／kg 0．9％氯化钠溶液1次）和NMDA受体拮抗剂组（70％中段小肠切除、腹腔注射1 mg／kg MK801溶液），每组再按术后1、3、5及7 d 分为4小组，每小组8只。每小组大鼠在术后对应时间点进行旷场试验后对海马组织进行取材，实时荧光定量聚合酶链反应检测大鼠海马NMDA受体1及犬尿氨酸氨基转移酶Ⅲ（KATⅢ）的mRNA表达；Western blot检测NMDA受体1的蛋白表达；高效液相-荧光法检测色氨酸（TRP）、犬尿氨酸（KYN）及犬尿喹啉酸（KYNA）的浓度；电镜观察海马神经元超微结构变化。结果术后1 d和3 d，与对照组比较，模型组大鼠运动得分减少（P＜0．05），休息时间、中央格子停留时间及外周／中央格子比均增加（P＜0．05）；NMDA受体1 mRNA及蛋白表达增加（P＜0．05）， KATⅢ mRNA表达减少（P＜0．05）；KYN／TRP及KYN／KYNA比值增加（P＜0．05）；拮抗剂组与模型组比较，术后1 d大鼠中央格子停留时间以及外周／中央格子比均降低（P＜0．05）；术后1 d和3 d NMDA受体1mRNA及蛋白表达降低（均为P＜0．05）。电镜结果显示，模型组海马神经元超微结构受损，拮抗剂组得到相应改善。结论 POFS大鼠海马NMDA受体表达增加，神经元存在一定程度的损伤，NMDA受体拮抗剂能改善POFS的中枢疲劳。Objective To explore the central mechanism of postoperative fatigue syndrome by detecting the expression of NMDA receptor and tryptophan metabolism. Methods After being numbered according to the weight, ninety-six male SD rats were randomly divided into control group （bowel loop was flipped after laparotomy and received intraperitoneal injection of saline at a dose of 1 ml/kg ）, POFS model （70% of the length of small intestine was resected and received intraperitoneal injection of saline at a dose of 1 ml/kg）, and NMDA antagonist groups （70% of the length of small intestine was resected and received intraperitoneal injection of MK801 at a dose of 1 ml/kg ）. Each group was divided into subgroups by postoperative 1, 3, 5 and 7 d, with 8 rats in each subgroup. The hippocampus was removed at each time point after open field test （OFT） to detect the mRNA expression levels of NMDA receptor 1 and kynurenine aminotransferase Ⅲ （KATⅢ） by real-time PCR. Protein level of NMDA receptor 1 was detected by Western blot. High performance liquid chromatography （HPLC） was used to measure the concentrations of tryptophan （TRP）, kynurenine （KYN） and kynurenic acid（KYNA）. Ultra-structural changes of hippocampal neurons were observed by transmission electron microscopy （TEM）.＆amp;nbsp;Results As compared to control group, exercise score decreased （P〈0.05）, rest time and central panel residence time prolonged, periphery/central panel ratio increased （all P〈0.05）, mRNA and protein expressions of NMDA receptor 1 increased （P〈0.05）, mRNA expression of KAT Ⅲ decreased （P〈0.05）, KYN/TRP ratio and KYN/KYNA ratio decreased （all P〈0.05） in POFS group on postoperative day 1 and 3. As compared to POFS group, central panel residence time and periphery/central panel ratio decreased on postoperative day 1 , and mRNA and protein expressions of NMDA receptor 1 decreased on postoperative day 1 and 3 （all P〈0.05） in antagonist group. TEM revealed that degenerated neuron

关 键 词：术后疲劳综合征 N-甲基-D-天冬氨酸受体 色氨酸 犬尿氨酸 大鼠 

分 类 号：R619[医药卫生—外科学]