本文選題：尿素通道蛋白B + 抑郁樣行為　； 參考：《重慶醫(yī)科大學》2013年碩士論文

【摘要】：目的：研究尿素通道蛋白B（Urea transporter B UT-B）基因敲除對小鼠自主行為的影響；以及小鼠海馬形態(tài)結構的改變，海馬組織內尿素和NO含量、MKP-1表達部位及水平的改變，以及MAPK信號分子的表達水平變化，探討UT-B基因敲除小鼠出現抑郁樣行為的分子機制。 方法：1.8-12周齡體重匹配，雌性野生型和UT-B基因敲除小鼠各20只，分別進行強迫游泳實驗，高架十字迷宮實驗和曠場實驗，，檢測野生型小鼠和UT-B基因敲除小鼠的自主行為。2.采用HE染色和免疫組織化學技術，觀察兩型小鼠海馬形態(tài)結構以及MKP-1的表達。3.采用尿素和NO檢測試劑盒測定兩型小鼠海馬尿素和NO水平。4.采用Western Blot技術檢測兩型海馬MKP-1、MAPKs相關信號分子的表達水平。 結果：1．強迫游泳實驗結果顯示：野生型小鼠和UT-B基因敲除小鼠的游泳不動時間分別為（86±13s）和（123±19s），UT-B基因敲除小鼠的游泳不動時間明顯延長（P=0.00120.05）。2．高架十字迷[笛榻峁允荊閡吧托∈蠛蚒T-B基因敲除小鼠開臂停滯時間分別為（74.79±13.70）s和（12.71±3.170）s（P=0.01090.05）；進入開臂次數分別為（12.50±1.645）和（4.75±0.56）（P=0.00210.05）；進入開臂次數與總入臂次數百分比分別為（58.63±4.28）%和（27.25±3.29）%（P=0.00090.05）；開臂停滯時間與總時間百分比分別為（31.61±6.71）%和（7.69±2.30）%（P=0.04260.05）。3．曠場實驗結果顯示：野生型小鼠和UT-B基因敲除小鼠在中央區(qū)域活動時間分別為（32.08±7.96）s和（13.50±2.96）s（P=0.0250.05）；中央區(qū)域活動路程分別為（873.5±137.1）mm和（594.8±190.8）mm（P=0.7780.05）；總路程分別為（10110±756.3）mm和（13770±844.7）mm（P=0.0250.05），UT-B基因敲除小鼠在中央區(qū)域活動時間明顯減少，運動頻率高。提示其具有抑郁樣行為。4．UT-B基因敲除小鼠海馬各亞區(qū)神經元密度減小，神經元MKP-1的表達水平上調。5．UT-B基因敲除和野生型小鼠海馬尿素水平為（16.7±2.0mM vs11.6±1.4mM，p=0.0040.001），NO含量為（1.05±0.07μmol/gprot vs1.29±0.06μmol/gprot，p=0.040.05），表明UT-B基因敲除小鼠海馬尿素水平顯著升高而NO含量明顯降低。6．UT-B基因敲除小鼠海馬MKP-1表達水平上調531.9%，pERK和pJNK表達水平明顯降低39.1%和21.3%，磷酸化p38MAPK水平無明顯改變。 結論：1．強迫游泳實驗、高架十字迷宮實驗和曠場實驗結果提示UT-B基因敲除小鼠具有抑郁樣行為。2．UT-B基因敲除引起小鼠海馬神經元減少，尿素堆積，NO含量減少。3．UT-B基因敲除引起小鼠海馬缺氧，誘導神經元MKP-1表達水平上調，MKP-1通過特異性下調pERK和pJNK水平引發(fā)小鼠的抑郁樣行為。
[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 鹵13 s and 123 鹵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 鹵7.96 s and 13.50 鹵2.96 s, respectively; the distance of central area activity was 0.873.5 鹵137.1)mm and 594.8 鹵190.8 UT-B / 0.7780.05, respectively; the total distance was 10110 鹵756.3)mm and 13770 鹵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 鹵2.0mM vs11.6 鹵1.4mmmmmMp0.0040.001. No content was 1.05 鹵0.07 渭 mol/gprot vs1.29 鹵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.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R749.41

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