MKP-1表达与UT-B基因敲除小鼠抑郁样行为的关系

发布时间：2018-04-18 22:02

本文选题：尿素通道蛋白B + 抑郁样行为　；参考：《重庆医科大学》2013年硕士论文

【摘要】：目的：研究尿素通道蛋白B（Urea transporter B UT-B）基因敲除对小鼠自主行为的影响；以及小鼠海马形态结构的改变，海马组织内尿素和NO含量、MKP-1表达部位及水平的改变，以及MAPK信号分子的表达水平变化，探讨UT-B基因敲除小鼠出现抑郁样行为的分子机制。方法：1.8-12周龄体重匹配，雌性野生型和UT-B基因敲除小鼠各20只，分别进行强迫游泳实验，高架十字迷宫实验和旷场实验，，检测野生型小鼠和UT-B基因敲除小鼠的自主行为。2.采用HE染色和免疫组织化学技术，观察两型小鼠海马形态结构以及MKP-1的表达。3.采用尿素和NO检测试剂盒测定两型小鼠海马尿素和NO水平。4.采用Western Blot技术检测两型海马MKP-1、MAPKs相关信号分子的表达水平。结果：1．强迫游泳实验结果显示：野生型小鼠和UT-B基因敲除小鼠的游泳不动时间分别为（86±13s）和（123±19s），UT-B基因敲除小鼠的游泳不动时间明显延长（P=0.00120.05）。2．高架十字迷[笛榻峁允荆阂吧托∈蠛蚒T-B基因敲除小鼠开臂停滞时间分别为（74.79±13.70）s和（12.71±3.170）s（P=0.01090.05）；进入开臂次数分别为（12.50±1.645）和（4.75±0.56）（P=0.00210.05）；进入开臂次数与总入臂次数百分比分别为（58.63±4.28）%和（27.25±3.29）%（P=0.00090.05）；开臂停滞时间与总时间百分比分别为（31.61±6.71）%和（7.69±2.30）%（P=0.04260.05）。3．旷场实验结果显示：野生型小鼠和UT-B基因敲除小鼠在中央区域活动时间分别为（32.08±7.96）s和（13.50±2.96）s（P=0.0250.05）；中央区域活动路程分别为（873.5±137.1）mm和（594.8±190.8）mm（P=0.7780.05）；总路程分别为（10110±756.3）mm和（13770±844.7）mm（P=0.0250.05），UT-B基因敲除小鼠在中央区域活动时间明显减少，运动频率高。提示其具有抑郁样行为。4．UT-B基因敲除小鼠海马各亚区神经元密度减小，神经元MKP-1的表达水平上调。5．UT-B基因敲除和野生型小鼠海马尿素水平为（16.7±2.0mM vs11.6±1.4mM，p=0.0040.001），NO含量为（1.05±0.07μmol/gprot vs1.29±0.06μmol/gprot，p=0.040.05），表明UT-B基因敲除小鼠海马尿素水平显著升高而NO含量明显降低。6．UT-B基因敲除小鼠海马MKP-1表达水平上调531.9%，pERK和pJNK表达水平明显降低39.1%和21.3%，磷酸化p38MAPK水平无明显改变。结论：1．强迫游泳实验、高架十字迷宫实验和旷场实验结果提示UT-B基因敲除小鼠具有抑郁样行为。2．UT-B基因敲除引起小鼠海马神经元减少，尿素堆积，NO含量减少。3．UT-B基因敲除引起小鼠海马缺氧，诱导神经元MKP-1表达水平上调，MKP-1通过特异性下调pERK和pJNK水平引发小鼠的抑郁样行为。
[Abstract]:Objective: to study the effect of urea channel protein B(Urea transporter B UT-B gene knockout on the autonomous behavior of mice, and the changes of hippocampal morphology, the expression of urea and no in hippocampus and the expression of MKP-1.To explore the molecular mechanism of depressive behavior in UT-B knockout mice.Methods Twenty female wild type and 20 UT-B knockout mice were used to perform forced swimming test, elevated cross maze test and open field test to detect the autonomous behavior of wild type mice and UT-B gene knockout mice.The hippocampal morphology and the expression of MKP-1 in the two types of mice were observed by HE staining and immunohistochemistry.The levels of urea and no in hippocampus of two type mice were measured by urea and no detection kit. 4. 4.Western Blot technique was used to detect the expression of MKP-1MAPKs related signal molecules in hippocampus.The result is 1: 1.The results of forced swimming test showed that the swimming immobility time of wild type mice and UT-B knockout mice was 86 卤13 s and 123 卤19 s respectively. The swimming immobility time of wild type mice and UT-B knockout mice was significantly prolonged.Elevated cross [?Fluffy knolls?Yeon Jing estranged?The results of open field test showed that the activity time of wild type mice and UT-B knockout mice in the central region were 32.08 卤7.96 s and 13.50 卤2.96 s, respectively; the distance of central area activity was 0.873.5 卤137.1)mm and 594.8 卤190.8 UT-B / 0.7780.05, respectively; the total distance was 10110 卤756.3)mm and 13770 卤844.7mm P0.0250.05 respectively.Domain activity time is significantly reduced,High frequency of motion.It is suggested that the density of neurons in the hippocampal subregions of UT-B knockout mice decreased.UT-B gene knockout and wild-type mice showed that the level of urea in hippocampus was 16.7 卤2.0mM vs11.6 卤1.4mmmmmMp0.0040.001. No content was 1.05 卤0.07 渭 mol/gprot vs1.29 卤0.06 渭 mol 路gprotptadine 0.040.05, which indicated that the level of urea in hippocampus of UT-B knockout mice was significantly higher than that of the control mice. 6.UT-B gene knockout mice had a significant decrease in no content. 6. UT-B gene knockout mice showed a significant increase in the level of urea in hippocampus and a significant decrease in the level of no in hippocampus of wild type mice.The expression of perk and pJNK in hippocampus decreased by 39.1% and 21.3%, but the level of phosphorylated p38MAPK did not change.Conclusion 1.The results of forced swimming test, elevated cross maze test and open field test showed that UT-B gene knockout mice had depressive behavior. 2. UT-B gene knockout induced the decrease of hippocampal neurons in mice.Decreased no content of UT-B gene knockout induced hypoxia in hippocampus of mice induced by UT-B gene knockout and up-regulated the expression of MKP-1 in neurons. MKP-1 specifically down-regulated the levels of pERK and pJNK and induced depressive behavior in mice.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R749.41

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