亚麻醉剂量氯胺酮介导NMDA-NO通路在小鼠术后认知功能障碍机制中的研究  

NMDA-NO pathway mediates the effect of a subanesthestic-dosage of ketamine on postoperative cognitive dysfunction in mice

作　　者：张锦曦 潘灵辉[1] Zhang Jinxi;Pan Linghui(Department of Anesthesiology,the Affiliated Tumor Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学附属肿瘤医院麻醉科,南宁530021

出　　处：《广西医科大学学报》2018年第5期652-656,共5页Journal of Guangxi Medical University

摘　　要：目的:研究亚麻醉剂量氯胺酮介导N-甲基-D-天冬氨酸受体(NMDA)-一氧化氮(NO)通路在小鼠术后认知功能障碍(POCD)中的作用机制。方法:将30只昆明小鼠随机分为空白组(A组)、手术组(B组)、手术+亚麻醉剂量氯胺酮组(C组),每组10只。A组不做任何处理,B、C两组行左颈动脉分离术,C组术中腹腔注射氯胺酮0.1 mg,3组小鼠于术后第1、第3、第5、第7天行水迷宫实验。术后9h取海马组织,采用免疫组化法检测NMDAR2B蛋白阳性表达情况,western blotting法检测NMDAR1、NMDAR2A、NMDAR2B蛋白表达,并检测海马组织一氧化氮合酶(NOS)活力及肿瘤坏死因子(TNF)-α、白介素(IL)-6的含量。结果:B组潜伏期在术后第1、第3天明显延长,目标象限百分比在术后第1、第3天明显降低(均P<0.05),A、C两组术后7d的潜伏期及目标象限百分比比较,差异均无统计学意义(均P>0.05)。NMDAR2B蛋白主要表达于海马CA1区,A组NMDAR2B蛋白表达呈中等阳性(++),B组NMDAR2B蛋白表达呈强阳性(+++),C组NMDAR2B蛋白表达呈弱阳性(+),B组免疫组化染色评分(IRS评分系统)显著高于A组和C组(P<0.05),而A组和C组比较差异无统计学意义(P>0.05)。B组海马组织NMDAR1、NMDAR2A、NMDAR2B蛋白表达、NOS活力及IL-6、TNF-α含量均较A组和C组显著升高(均P<0.05),而A组和C组比较差异无统计学意义(P>0.05)。结论:亚麻醉剂量氯胺酮能拮抗NMDA受体蛋白表达,抑制NOS活性,减少信号通路下游神经炎症因子释放,从而有效抑制小鼠术后POCD。Objective：To study the regulation of NMDA-NO pathway on postoperative cognitive dysfunction（POCD）in mice treated by a subanesthestic-dosage of ketamine.Methods：Thirty mice were randomly divided into three groups（n=10 per group）：blank control group（group A）,surgery group（group B）,and surgery plus ketamine group（group C）.The mice in group B and C were subjected to the separation of left carotid artery.Mice in group C were intraperitoneally injected with a subanesthetic dose of ketamine（0.1 mg）during the surgery.The water maze test was performed on the 1 st,3 rd,5 th and 7 th day after operation.The hippocampus were taken at 9 hpost-operation.The protein expression of N-methyl-D-aspartic acid receptor（NMDAR）2B in hippocampus were determined by immunohistochemistry.NMDAR1,NMDAR2A,and NMDAR2B protein levels were detected by western blotting.The contents of interleukins（IL-6）and tumor necrosis factor（TNF-α）in hippocampus and the relative activity of nitric oxide synthase（NOS）were measured.Results：After 1 and 3 days of operation,the latency to find the platform of the group B was significantly increased,while percent time in the target quadrant of water maze was decreased.There was no significant difference in the latency and percent time in the target quadrant between group A and C（P 〉0.05）.NMDAR2B located on CA1 region of hippocampus.It was strongly expressed in group B,moderately expressed in group A,and weakly expressed in group C.The IRS score,the protein expressions of NMDAR1,NMDAR2A and NMDAR2B as well as the relative activity of NOS in group B were significantly higher than those in group A and C（P 〈0.05）,but no significant difference was noted between group A and C（P〉 0.05）.Conclusion：A subanesthestic-dosage of ketamine could antagonize NMDA receptors,inhibit the activity of NOS and reduce the release of inflammatory factors,thereby reducing the incidence of POCD.

关 键 词：亚麻醉剂量氯胺酮 术后认知功能障碍 N-甲基-D-天冬氨酸受体 炎症因子 

分 类 号：R749.2[医药卫生—神经病学与精神病学]