本文關(guān)鍵詞： HIBD動(dòng)物模型 抑癌蛋白PTEN 時(shí)間依賴性 多肽Tat-K13劑量 PTEN泛素化 行為學(xué)變化 學(xué)習(xí)記憶　出處：《重慶醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：第一部分HIBD動(dòng)物模型的建立和PTEN入核水平的變化 目的：為了建立新生鼠缺氧缺血性腦損傷（HIBD）實(shí)驗(yàn)動(dòng)物模型，采用7日齡SD新生鼠，觀察抑癌蛋白PTEN是否在新生鼠缺氧缺血后入核增加。 方法：新生7日齡SD大鼠，隨機(jī)分成HIBD組、假手術(shù)組。用異氟烷進(jìn)行麻醉，仰臥固定于手術(shù)板上，75%酒精消毒頸部皮膚，偏左剪開，輕輕分離皮下脂肪組織，找到左側(cè)頸總動(dòng)脈后用4-0絲線結(jié)扎兩端并剪短血管，檢查斷流成功后，縫合切口。假手術(shù)組分離左側(cè)頸總動(dòng)脈后不結(jié)扎，縫合切口。術(shù)后放回母鼠身邊休息兩小時(shí)，HIBD組放入密閉缺氧箱，持續(xù)充以8%氧和92%氮混合氣，持續(xù)2.5小時(shí)，取出放回母鼠身邊飼養(yǎng)。假手術(shù)組不做缺氧。造模后分別于缺氧3h、6h、12h、24h、3d、7d，冰上分離左右側(cè)海馬和皮層，提取左側(cè)海馬核蛋白，用western blot檢測(cè)核蛋白中PTEN水平的變化。 結(jié)果：新生鼠缺氧缺血性腦損傷后3h、6h、12h， PTEN入核水平較假手術(shù)組高，24h后幾乎恢復(fù)到假手術(shù)水平。 結(jié)論：新生鼠缺氧缺血時(shí)，PTEN入核水平先增加后恢復(fù)正常，兩者之間存在時(shí)效關(guān)系，，找到這種關(guān)系有利于我們找到最佳治療時(shí)間窗。 第二部分：多肽Tat-K13治療HIBD后的動(dòng)物行為學(xué)檢測(cè) 目的：在第部分實(shí)驗(yàn)結(jié)果的基礎(chǔ)上，檢測(cè)多肽Tat-K13對(duì)抑制PTEN入核的劑效關(guān)系。應(yīng)用多種動(dòng)物行為學(xué)檢測(cè)方法，觀察Tat-K13多肽對(duì)HIBD的治療作用。 方法：首先在完成模型的新生鼠中隨機(jī)分組，腹腔注射不同劑量的Tat-K13多肽（0、2.5、5、10、20mg/kg）或Tat-K13R對(duì)照多肽，于缺氧前0h和缺氧3h各給次藥，在缺氧12h時(shí)取腦組織，提左側(cè)海馬組織核蛋白，western blot檢測(cè)PTEN入核水平的變化。劑效關(guān)系穩(wěn)定后，在動(dòng)物模型上給藥，分別于缺氧0h、3h、24h、48h各腹腔注射次多肽。4-6周后觀察行為學(xué)變化。分別做了體重測(cè)量、高架十字迷宮、轉(zhuǎn)棒測(cè)試、抓力測(cè)試、水迷宮測(cè)試和避暗逃避實(shí)驗(yàn)。 結(jié)果：根據(jù)劑效關(guān)系圖，確定用藥劑量為10mg/kg。體重在四個(gè)實(shí)驗(yàn)組中沒有統(tǒng)計(jì)學(xué)差異；高架十字迷宮沒有統(tǒng)計(jì)學(xué)差異，說明HIBD不造成新生鼠的情感記憶障礙；轉(zhuǎn)棒實(shí)驗(yàn)，HIBD組較Sham組在棒上的時(shí)間長，給予多肽K13治療能部分逆轉(zhuǎn)。抓力實(shí)驗(yàn)，HIBD組右側(cè)前肢肌力較左側(cè)強(qiáng)，多肽K13能部分逆轉(zhuǎn)，基本達(dá)Sham組水平；水迷宮實(shí)驗(yàn)，檢測(cè)其空間記憶能力，HIBD組較Sham組找到平臺(tái)的時(shí)間長，多肽K13組較HIBD組找到平臺(tái)的時(shí)間長。避暗逃避實(shí)驗(yàn)主要也檢測(cè)其空間記憶力，HIBD組較Sham組進(jìn)入電擊箱的潛伏期短，K13組較HIBD組潛伏期幾乎達(dá)Sham組水平。說明HIBD對(duì)空間記憶能力有損害作用，多肽K13能部分逆轉(zhuǎn)這種損害。 結(jié)論：抑制PTEN入核對(duì)新生鼠缺氧缺血性腦損傷有定的治療作用。
[Abstract]:Part I Establishment of HIBD animal model and changes of PTEN entry level. Aim: to establish an experimental model of hypoxic-ischemic brain injury (HIBD) in neonatal rats, 7-day-old SD neonatal rats were used to observe whether the tumor suppressor protein PTEN increased after hypoxic ischemia in neonatal rats. Methods: SD rats of 7 days old were randomly divided into HIBD group and sham operation group. Anesthetized with isoflurane, 75% alcohol was fixed on the surgical plate to sterilize the neck skin. The skin was cut to the left and subcutaneous adipose tissue was gently separated. After finding the left common carotid artery, the left common carotid artery was ligated with 4-0 silk thread and the blood vessels were cut off. After the disconnection was successful, the incision was sutured. The sham operation group did not ligate the left common carotid artery after separating the left common carotid artery. Suture incision. After the operation, the rats were put back to the mother for two hours to rest. In the HIBD group, the rats were placed in a closed hypoxia chamber, filled with 8% oxygen and 92% nitrogen mixture for 2.5 hours. The sham-operation group did not do anoxia. After 3 h of hypoxia, the left and right hippocampus and cortex were isolated from the left and right hippocampus and cortex for 7 days. The changes of PTEN level in the left hippocampus were detected by western blot. Results: the level of PTEN entry in neonatal rats was higher than that in sham-operated group for 24 h and almost recovered to the level of sham-operation after 3 hours and 12 hours after hypoxic-ischemic brain injury. Conclusion: the nuclear entry level of PTEN in neonatal rats increased first and then returned to normal, and there was a time-dependent relationship between the two. Finding this relationship is helpful for us to find the best therapeutic time window. The second part: the detection of animal behavior after polypeptide Tat-K13 treatment of HIBD. Aim: based on the results of the first part of the experiment, the effects of polypeptide Tat-K13 on inhibiting the entry of PTEN into the nucleus were detected. The therapeutic effect of Tat-K13 polypeptide on HIBD was observed by various animal behavioral methods. Methods: first of all, the neonate rats were randomly divided into two groups. The rats were injected intraperitoneally with different doses of Tat-K13 polypeptide (0 2. 5%) or Tat-K13R control peptide (10 mg / kg). The brain tissues were taken at 12 h after hypoxia, 0 h before hypoxia and 3 h after hypoxia. Western blot was extracted from the left hippocampal tissue to detect the changes of PTEN entry level. After stable dose-effect relationship, the rats were given the drug on the animal model, and the behavioral changes were observed after 48h intraperitoneal injection of polypeptide at 3h, 24h or 48h, respectively, and the body weight was measured. Elevated cross maze, rotating bar test, grip test, water maze test and escape test. Results: according to the dose-effect diagram, the dosage was determined to be 10 mg / kg. There was no significant difference in body weight among the four experimental groups, and there was no statistical difference in the elevated cross maze, which indicated that HIBD did not cause emotional memory disorder in newborn rats. HIBD group had a longer time on the stick than Sham group, and the treatment of peptide K13 could be partially reversed. The muscle strength of right forelimb was stronger than that of left side in the grip test group, and the peptide K13 could be partially reversed to the level of Sham group, and the water labyrinth test showed that the muscle strength of right forelimb in HIBD group was stronger than that in left side. The spatial memory ability of HIBD group was longer than that of Sham group. The time of finding platform in K13 group was longer than that in HIBD group, and the spatial memory in HIBD group was shorter than that in Sham group. The incubation period of K13 group was almost equal to that of Sham group compared with HIBD group. Memory is impaired, Peptide K 13 can partially reverse this damage. Conclusion: inhibition of PTEN entry nucleus has definite therapeutic effect on hypoxic ischemic brain injury in neonatal rats.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R742

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張鵬;程國強(qiáng);;亞低溫治療新生兒缺氧缺血性腦病的研究進(jìn)展[J];中國當(dāng)代兒科雜志;2013年10期

2 李虎;萬俊;張玲;;亞低溫、川芎嗪對(duì)缺氧缺血性腦病幼鼠腦組織自由基代謝的影響[J];兒科藥學(xué)雜志;2013年09期

3 李虎;萬俊;張玲;;亞低溫川芎嗪對(duì)缺氧缺血性腦病幼鼠的腦保護(hù)作用[J];安徽醫(yī)學(xué);2013年10期

4 王仁君;蘇紀(jì)平;;膜受體內(nèi)化的途徑及其相關(guān)疾病的研究進(jìn)展[J];廣東醫(yī)學(xué);2013年20期

5 曹長青;李宇寧;楊學(xué)梅;弓毅谷;王芳;李衛(wèi)國;;亞低溫治療新生兒缺氧缺血性腦病遠(yuǎn)期臨床效果的Meta分析[J];中國當(dāng)代兒科雜志;2015年02期

6 潘玲麗;屈藝;羅黎力;趙靜;李姣;唐軍;伍金林;李熙鴻;母得志;;整合素連接激酶在新生鼠缺氧缺血性腦損傷中的表達(dá)及作用[J];臨床兒科雜志;2014年08期

7 楊坦;王衛(wèi)娜;;川芎嗪對(duì)d-半乳糖所致衰老大鼠學(xué)習(xí)記憶功能和海馬CA1區(qū)長時(shí)程增強(qiáng)的影響[J];中國民族民間醫(yī)藥;2015年01期

8 Xianchao Li;Wensheng Hou;Xiaoying Wu;Wei Jiang;Haiyan Chen;Nong Xiao;Ping Zhou;;660 nm red light-enhanced bone marrow mesenchymal stem cell transplantation for hypoxic-ischemic brain damage treatment[J];Neural Regeneration Research;2014年03期

9 沈陽;陳宇;傅潔瑜;葉玉如;;老年癡呆癥的分子機(jī)制[J];生命科學(xué);2014年06期

10 周啟心;毛榕榕;徐林;;令人煩惱的記憶[J];生命科學(xué);2014年06期

相關(guān)博士學(xué)位論文 前10條

1 楊文建;金針菇多糖的分離純化、結(jié)構(gòu)分析及其記憶功能改善作用機(jī)制研究[D];南京農(nóng)業(yè)大學(xué);2012年

2 譚濤;低頻rTMS增強(qiáng)Aβ模型大鼠學(xué)習(xí)記憶功能的研究[D];天津醫(yī)科大學(xué);2013年

3 汪之沫;SRF-IEG通路在束縛應(yīng)激所致內(nèi)臟高敏感中的作用研究[D];華中科技大學(xué);2013年

4 柴高尚;下調(diào)pp32恢復(fù)p300/CBP組蛋白乙�；D(zhuǎn)移酶活性改善學(xué)習(xí)記憶障礙[D];華中科技大學(xué);2013年

5 劉羽;HDAC2對(duì)APP/PS1轉(zhuǎn)基因阿爾茨海默病模型小鼠的作用及機(jī)制研究[D];北京協(xié)和醫(yī)學(xué)院;2013年

6 張峗;TLR2在骨髓間充質(zhì)干細(xì)胞修復(fù)缺氧缺血性腦損傷中的作用及機(jī)制[D];重慶醫(yī)科大學(xué);2013年

7 朱槞槞;頻率和時(shí)間模式依賴的海馬突觸可塑性及其機(jī)制研究[D];中國科學(xué)技術(shù)大學(xué);2012年

8 牟海燕;發(fā)育期大鼠高級(jí)視皮層活動(dòng)對(duì)初級(jí)視皮層突觸可塑性影響的研究[D];天津醫(yī)科大學(xué);2010年

9 王華龍;重復(fù)經(jīng)顱磁刺激改善老化相關(guān)的認(rèn)知功能損傷的電生理機(jī)制及潛在代謝產(chǎn)物的變化[D];河北醫(yī)科大學(xué);2014年

10 王震;ICA69通過C末端結(jié)構(gòu)域影響PICK1-PKCα復(fù)合物運(yùn)輸和小腦可塑性[D];浙江大學(xué);2014年

相關(guān)碩士學(xué)位論文 前10條

1 張曉燕;甘氨酸誘導(dǎo)的NMDA受體電流的雙向可塑性[D];南京醫(yī)科大學(xué);2013年

2 吳玨;睡眠與陳述性記憶鞏固[D];華東師范大學(xué);2013年

3 柴寧;去甲腎上腺素促進(jìn)恐懼記憶消退的長期維持[D];河北醫(yī)科大學(xué);2013年

4 朱靜芳;恐懼關(guān)聯(lián)特征對(duì)恐懼泛化及恐懼消退過程的影響[D];西南大學(xué);2013年

5 闞宇;慢性痛大鼠海馬NO/cGMP/PKG信號(hào)通路在針刺累積鎮(zhèn)痛效應(yīng)中的作用分析[D];中國中醫(yī)科學(xué)院;2013年

6 秦秋紅;東方蜜蜂與西方蜜蜂學(xué)習(xí)記憶比較及蜜蜂學(xué)習(xí)記憶相關(guān)分子機(jī)理分析[D];江西農(nóng)業(yè)大學(xué);2013年

7 楊遙;REM睡眠剝奪對(duì)小鼠海馬tau蛋白磷酸化的影響[D];石河子大學(xué);2013年

8 陳善平;星型膠質(zhì)細(xì)胞來源的ATP對(duì)海馬CA1區(qū)的LTD具有重要的調(diào)控作用[D];南方醫(yī)科大學(xué);2013年

9 胡瀟方;磷酸化AKT在乳腺癌組織中的表達(dá)及其對(duì)淋巴轉(zhuǎn)移的作用[D];泰山醫(yī)學(xué)院;2013年

10 劉瑩;AMPA受體在亞慢性染鋁致大鼠學(xué)習(xí)記憶損傷中的作用[D];山西醫(yī)科大學(xué);2013年

本文編號(hào)：1510622