本文選題：海馬齒狀回　切入點：轉(zhuǎn)位蛋白(TSPO)　出處：《河北北方學(xué)院》2017年碩士論文

【摘要】：18k Da轉(zhuǎn)位蛋白(18k Da translocator protein,TSPO)在中樞神經(jīng)系統(tǒng)中,TSPO主要分布在膠質(zhì)細胞線粒體外膜上,能夠介導(dǎo)膽固醇從線粒體外膜向線粒體內(nèi)轉(zhuǎn)運,參與神經(jīng)類固醇合成的限速步驟,介導(dǎo)神經(jīng)類固醇合成尤其是四氫孕酮的合成,而神經(jīng)類固醇作為神經(jīng)調(diào)節(jié)遞質(zhì),影響γ-氨基丁酸,谷氨酸受體、等多個神經(jīng)遞質(zhì)及其受體功能,在應(yīng)激反應(yīng)或焦慮,抑郁,創(chuàng)傷后應(yīng)激障礙等應(yīng)激相關(guān)疾病中起到重要的調(diào)節(jié)作用。近年來研究發(fā)現(xiàn),TSPO可能成為情感性精神疾病尤其是焦慮癥抑郁癥治療的潛在靶標。本研究旨在探究小鼠海馬齒狀回18k Da轉(zhuǎn)位蛋白(TSPO)過表達對焦慮,抑郁樣行為的影響,為TSPO成為藥物潛在新靶標提供實驗依據(jù)。本研究成年雄性ICR小鼠海馬齒狀回微注射攜帶TSPO基因的慢病毒,以致TSPO過表達,觀察小鼠海馬齒狀回區(qū)注射慢病毒后注射位置熒光蛋白表達準確性以及TSPO蛋白表達水平的變化;第12,14和16天進行小鼠高架十字迷宮、爬梯實驗、明暗箱實驗檢測焦慮樣行為,第18,和21天進行小鼠懸尾和強迫游泳兩個行為絕望模型檢測抑郁樣行為;同時采用Western Blot和ELISA方法分別檢測海馬注射孔周圍3mm組織內(nèi)TSPO蛋白水平以及四氫孕酮含量,初步探討海馬齒狀回過表達TSPO與焦慮,抑郁障礙的關(guān)系。本研究結(jié)果如下:慢病毒定位注射后12天,過表達組小鼠海馬組織內(nèi)TSPO蛋白水平明顯高于空白組(P=0.001);注射后12天,過表達組小鼠海馬齒狀回區(qū)綠色熒光蛋白明顯;小鼠海馬齒狀回微注射慢病毒后12天,對小鼠自發(fā)活動無顯著影響(P0.05)。與LV-NC組相比,LV-TSPO組顯著增加小鼠進入中心區(qū)時間(P0.05);顯著增加小鼠進入開臂次數(shù)和開臂時間的百分比(P0.05;P0.01);顯著減少小鼠站立次數(shù)(P0.05);顯著增加小鼠進入明箱的次數(shù)和時間(P0.05;P0.01),提示海馬齒狀回TSPO過表達小鼠表現(xiàn)出顯著的抗焦慮樣行為作用;與LV-NC組相比,LV-TSPO組明顯減少小鼠在懸尾實驗和強迫游泳實驗中的不動時間(P0.001;P0.05),提示海馬齒狀回TSPO過表達小鼠表現(xiàn)出顯著的抗抑郁樣行為作用。進一步的生化檢測表明,與陰性對照組相比,小鼠海馬齒狀回TSPO表達可導(dǎo)致海馬注射孔周圍3mm組織的TSPO蛋白表達水平顯著增加(P0.05),并且其四氫孕酮含量顯著增加(P0.05)。小鼠海馬齒狀回過表達TSPO具有顯著的抗焦慮、抗抑郁樣的行為學(xué)效應(yīng),且TSPO介導(dǎo)的四氫孕酮水平升高可能是其重要機制。
[Abstract]:In the central nervous system (CNS), the 18kDa translocation protein (18kDa translocator) mainly distributes on the mitochondrial outer membrane of glial cells, which can mediate the transport of cholesterol from the mitochondrial outer membrane to the mitochondria, and participate in the rate-limiting steps of neurosteroid synthesis. Mediates the synthesis of neurosteroids, especially tetrahydroprogesterone, and neurosteroids, as neuroregulatory transmitters, affect the functions of many neurotransmitters, such as 緯 -aminobutyric acid, glutamate receptors, and their receptors, in stress response or anxiety, depression, etc. In recent years, it has been found that TSPO may be a potential target for the treatment of affective mental disorders, especially anxiety disorders. Overexpression of 18kDa translocation protein TSPO in dentate gyrus of sea horse, The effect of depressive behavior provides experimental evidence for TSPO to become a potential new target for drugs. In this study, the adult male ICR mice were microinjected with lentivirus carrying TSPO gene in dentate gyrus, which resulted in overexpression of TSPO. The accuracy of fluorescent protein expression and the expression level of TSPO protein were observed after injecting lentivirus into the dentate gyrus of mice. On the 18th and 21st days, the despairing model of suspending tail and forced swimming in mice was used to detect depressive behavior, and the TSPO protein level and tetrahydroprogesterone content in the 3mm tissue around the hippocampal injection pore were detected by Western Blot and ELISA, respectively. To explore the relationship between the expression of TSPO and anxiety and depression in dentate gyrus of the sea horse. The results are as follows: 12 days after lentivirus injection, the level of TSPO protein in hippocampus of the overexpression group was significantly higher than that of the control group, and 12 days after the injection, the expression of TSPO protein in the hippocampus of the control group was significantly higher than that of the control group. The green fluorescent protein in the dentate gyrus of the sea horse was obvious in the overexpression group, and 12 days after the microinjection of lentivirus in the dentate gyrus of the mice. Compared with the LV-NC group, the LV-TSPO group significantly increased the entry time of mice into the central area, significantly increased the number of times of entering the open arm and the percentage of the opening arm time of the mice, and significantly decreased the standing times of the mice (P0.05). The times and time of entering the box indicated that the over-expression of TSPO in the dentate gyrus of the sea horse showed significant anti-anxiety behavior in mice. Compared with LV-NC group, LV-TSPO group significantly reduced the immobility time of mice in tail suspension test and forced swimming test, suggesting that TSPO overexpression in dentate gyrus of sea horse showed significant antidepressant behavior. Compared with the negative control group, The expression of TSPO in the dentate gyrus of the sea horse significantly increased the expression of TSPO protein in the 3mm tissue around the hippocampal injection pore, and the content of tetrahydroprogesterone increased significantly. The expression of TSPO in the dentate gyrus of the mice was significantly anti-anxiety. The behavioral effect of antidepressant and the elevated level of tetrahydroprogesterone mediated by TSPO may be the important mechanism.
【學(xué)位授予單位】：河北北方學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R749

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