复合型肾虚质大鼠神经行为障碍与NMDA受体表达研究

孙瑜嬬等

摘要：目的 探討肾虚质与脑功能衰退的相关性及肾藏志理论的物质基础与神经生物学机制。方法 采用“猫吓鼠”造模法造成先天不足加后天失养复合型肾虚模型，随机分为模型组、左归丸组和右归丸组；空白组产自正常孕鼠。对仔鼠恐吓同时开始灌胃，左归丸组、右归丸组分别给予左归丸、右归丸混悬液，空白组和模型组给予等量生理盐水，1次/d，连续2个月。悬吊试验和Y迷宫实验观测大鼠运动和空间学习记忆能力，免疫组化法检测N-甲基-D-天冬氨酸（NMDA）受体亚基NR2A、NR2B蛋白表达。结果 悬吊试验：模型组持续时间低于空白组（P<0.05），左归丸组和右归丸组高于模型组（P<0.05）；Y迷宫实验：模型组跑至安全区正确次数低于空白组（P<0.05），左归丸组和右归丸组高于模型组（P<0.05）；模型组NR2A、NR2B光密度值较空白组降低（P<0.05），左归丸组和右归丸组光密度值较模型组升高（P<0.05）。结论 肾虚质大鼠神经行为学异常且海马NMDA受体亚基表达降低；左归丸、右归丸可上调NMDA受体表达且改善大鼠脑功能，揭示了肾虚质与脑功能衰退的相关性及肾藏志理论的物质基础与神经生物学机制。

关键词：肾虚质；悬吊试验；Y迷宫实验；NR2A蛋白；NR2B蛋白；大鼠

DOI：10.3969/j.issn.1005-5304.2015.09.015

中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2015）09-0052-04

Study on Neurobehavioral Disorders and NMDA Receptor Expression in Rats with Kidney-deficiency Constitution SUN Yu-ru， SUN Yao-guang， ZHANG Qi， WANG Xiao-di， WANG Xing， SUN Li-jun （Shaanxi University of Chinese Medicine， Xianyang 712046， China）

Abstract：Objective To discuss the correlation between kidney-deficiency constitution and brain function decline， and material basis of kidney-storing-spirit theory and neurobiology mechanism. Methods By employing the method of “cats scare rats”， composite offspring rat models with deficiency and acquired dystrophy were built， then they were divided into model group， Zuogui Pill group and Yougui Pill group. Blank group was composed from normal pregnant rats. When the intimidation of offspring began， Zuogui Pill and Yougui Pill groups received gavage with corresponding doses of medicine， 1 times per day for 2 consecutive months. Model and blank groups received the same amount of normal saline. Exercise capacity was detected by suspension test. Learning and memory capacity was detected by Y maze test. Expressions of NMDA receptor subunits NR2A and NR2B were detected by immunohistochemical method. Results In suspension test， the duration of model group was shorter than blank group （P<0.05）， while duration of Zuogui Pill and Yougui Pill groups was longer than the model group （P<0.05）. In Y maze test， the correct number of model group was less than blank group， and increased significantly after treatment （P<0.05）. The optical density of NR2A and NR2B was lower in model group than blank group （P<0.05） and higher in Zuogui Pill and Yougui Pill groups than model group （P<0.05）. Conclusion The neurobehavioral is abnormal and NMDA receptor expression depresses in rats with kidney- deficiency constitution. Zuogui Pill and Yougui Pill can rise NMDA receptor expression and improve brain function of rats， which reveal the correlation between kidney-deficiency constitution and brain function decline， and material basis of kidney-storing-spirit theory and neurobiology mechanism.

Key words：kidney-deficiency constitution；suspension test；Y maze test；NR2A；NR2B；rats

基金项目：陕西省科技厅研究项目（2013JM4033）；陕西省重点实验室研究项目（13JS031）

《灵枢·本神》曰：“恐惧而不解则伤精，精伤者则骨酸痿厥，精时自下。”王冰注释：“恐而不已则内感于肾，故伤也。”中医理论认为，肾为先天之本，藏有先天之精和后天之精，肾精的盛衰决定着子代禀赋的强弱。肾藏精生髓，脑为髓海，主神志思维，司机体运动。髓由肾精及气血所化，充养脑实质，肾虚则经不上承而脑髓空虚、脑窍失养致脑失其用[1]。本实验以肾虚质大鼠运动、学习等神经行为学改变和海马N-甲基-D-天冬氨酸（NMDA）受体亚基NR2A、NR2B蛋白表达为切入点，探讨肾虚质和脑功能衰退的相关性，以及肾藏志理论的物质基础和神经生物学机制。

1 实验材料

1.1 动物

8周龄清洁级SD大鼠40只，雌雄各半，体质量200～300 g，西安交通大学医学院实验动物中心，许可证号SCXK（陕）2007-001。……