GLYX-13通过NR2B-ERK-CREB信号通路改善氯胺酮麻醉后幼鼠认知功能障碍

GLYX-13 improves cognitive dysfunction in young rats after ketamine anesthesia through NR2B-ERK-CREB signaling pathway

目的:探讨GLYX-13通过N-甲基-D-门冬氨酸受体2B(NR2B)-胞外信号调节激酶(ERK)-环磷腺苷效应元件结合蛋白(CREB)信号通路改善氯胺酮麻醉后幼鼠认知功能障碍。方法:将60只幼鼠根据随机数字法分为对照组(NS组)、氯胺酮麻醉组(Ket组)、氯胺酮麻醉+GLYX-13组(Ket+GLYX-13组),每组20只。Ket组和Ket+GLYX-13组小鼠每天腹腔注射30 mg/kg氯胺酮,间隔30 min重复一次,每天3次;Ket+GLYX-13组小鼠每天首次注射氯胺酮前2 h尾静脉注射1 mg/kg GLYX-13;NS组每天注射等量生理盐水,共5 d。水迷宫实验和旷场实验测定小鼠认知功能,Western blot法测定海马组织NR2B、ERK、磷酸化ERK(p-ERK)、CREB、p-CREB蛋白水平,免疫荧光法测定海马CA1区NR2B、p-ERK、p-CREB蛋白表达情况。结果:与NS组比较,Ket组和Ket+GLYX-13组小鼠逃避潜伏期延长(P<0.05),穿越原平台象限次数和在原平台象限停留时间降低(P<0.05),中心区停留时间均缩短(P<0.05),海马组织NR2B、p-ERK、p-CREB蛋白水平降低(P<0.05),海马CA1区NR2B、p-ERK、p-CREB荧光阳性信号强度降低(P<0.05)。与Ket组比较,Ket+GLYX-13组小鼠逃避潜伏期缩短(P<0.05),穿越原平台象限次数和在原平台象限停留时间升高(P<0.05),中心区停留时间均延长(P<0.05),海马组织NR2B、p-ERK、p-CREB蛋白水平升高(P<0.05),海马CA1区NR2B、p-ERK、p-CREB荧光阳性信号强度升高(P<0.05)。结论:GLYX-13通过激活NR2B-ERK-CREB信号通路改善氯胺酮麻醉后幼鼠认知功能障碍。

Objective:To investigate the effect of GLYX-13 on cognitive dysfunction in young rats after ketamine anesthesia through N-methyl-D-aspartate receptor 2 B(NR2 B)-extracellular signal-regulated kinase(ERK)-cyclic adenosine response element binding protein(CREB)signaling pathway.Methods:Sixty young rats were divided into control group(NS group),ketamine anesthesia group(Ket group)and ketamine anesthesia+GLYX-13 group(Ket+GLYX-13 group)accorded to random number method,20 rats in each group.The Ket group and Ket+GLYX-13 group mice were intraperitoneally injected with 30 mg/kg ketamine,repeat at intervals of 30 min,3 times a day;the Ket+GLYX-13 group mice were injected with 1 mg/kg GLYX-13 intravenously 2 h before the first injection of ketamine per day;the NS group mice were injected with the same amount of normal saline daily.All for 5 days.The water maze test and open field experiment were used to determine the cognitive function of mice.Western blot analysis was used to determine the NR2 B,ERK,phosphorylated ERK(p-ERK),CREB and p-CREB protein levels in hippocampus.The expression of NR2 B,p-ERK and p-CREB protein in hippocampal CA1 region were determined by immunofluorescence.Results:Compared with the NS group,the escape latency was prolonged(P<0.05),the number of quadrants crossing the original platform and the residence time of the original platform quadrant were decreased(P<0.05),the residence time of the central region was shortened(P<0.05),the levels of NR2 B,p-ERK and p-CREB protein in hippocampus were decreased(P<0.05),the positive signal intensity of NR2 B,p-ERK and p-CREB in hippocampal CA1 region was decreased(P<0.05)of the Ket group and the Ket+GLYX-13 group.Compared with Ket group,the escape latency was shortened(P<0.05),the number of quadrants crossing the original platform and the residence time in the original platform quadrant were increased(P<0.05),the residence time in the central area was prolonged(P<0.05),the levels of NR2 B,p-ERK and p-CREB proteins in hippocampus were increased(P<0.05),the positive signal intensity of NR2 B,p-ERK and p-CREB in hippocampal CA1 region were increased(P<0.05)of mice the Ket+GLYX-13 group.Conclusion:GLYX-13 can improve the cognitive dysfunction of young rats after ketamine anesthesia by activating the NR2 B-ERK-CREB signaling pathway.