本文選題：雷帕霉素 + mTOR信號通路�。� 參考：《浙江大學(xué)》2014年碩士論文

【摘要】：目的：紅藻氨酸(KA)誘導(dǎo)的癲癇發(fā)作可激活哺乳動物的mammalian target of rapamycin (mTOR)信號通路作用靶點使下游S6蛋白磷酸化增加,mTOR抑制劑雷帕霉素可以抑制mTOR的激活,且可能有后續(xù)的潛在抗癲癇作用。然而,初步研究表明在KA注射前短時間內(nèi)給予雷帕霉素,S6磷酸化反而引起mTOR信號通路的激活。在本研究中,我們同時在正常大鼠和KA癲癇模型大鼠進行了更詳細(xì)地研究,探討雷帕霉素這一矛盾性作用的規(guī)律。 方法：正常組大鼠或KA模型組大鼠提前在不同時間點給予雷帕霉素,在給予雷帕霉素或癲癇發(fā)作后的不同時間點(1,3,6,10,15和24h)處死提取腦組織蛋白,采用Western Blotting(?)對mTOR信號靶點Akt, mTOR, Rictor, Raptor, S6K和S6的蛋白磷酸化進行了分析。對癲癇發(fā)作行為活動進行行為學(xué)監(jiān)測,并根據(jù)改良的Racine法進行分級(每個時間點n=6)。使用Fluoro-Jade B染色檢測神經(jīng)細(xì)胞的死亡 結(jié)果：我們發(fā)現(xiàn)在正常大鼠中,在注射雷帕霉素的3h-24h后,其表現(xiàn)出預(yù)期的對S6磷酸化的劑量依賴性抑制,然而在給予雷帕霉素后1h則發(fā)現(xiàn)了S6磷酸化水平反常的升高。在KA造模組大鼠中,在造模前10h以上使用雷帕霉素進行預(yù)處理可抑制KA癲癇引起的mTOR激活,相比之下,在KA造模前1h-6h使用雷帕霉素則會反常地加劇KA癲癇引起的mTOR激活。與對照組相比,在KA造模前1h使用雷帕霉素進行預(yù)處理的大鼠,癲癇發(fā)作更嚴(yán)重、持續(xù)時間更長,且神經(jīng)細(xì)胞死亡數(shù)增加。而在KA造模前10h以上使用雷帕霉素進行預(yù)處理則對癲癇沒有影響且能減少神經(jīng)細(xì)胞的死亡。雷帕霉素對S6磷酸化的這種反常效應(yīng)與上游的mTOR信號的改變相一致,且這種效應(yīng)可被哌立福辛(一種Akt抑制劑)逆轉(zhuǎn)。 意義：這些數(shù)據(jù)表明了雷帕霉素對S6蛋白磷酸化的調(diào)節(jié)及其對癲癇發(fā)作的復(fù)雜性作用。在臨床應(yīng)用雷帕霉素作為癲癇及其他神經(jīng)內(nèi)科的治療用藥中應(yīng)充分考慮到該反常作用。
[Abstract]:Aim: epileptic seizures induced by kainic acid can activate the mammalian target of rapamycin mTOR-signaling pathway in mammals, and the downstream phosphorylation of S6 protein can be increased by rapamycin, a inhibitor of kainic acid, which can inhibit the activation of mTOR. And may have subsequent potential antiepileptic effects. However, preliminary studies have shown that the phosphorylation of rapamycin I S6 at short time before Ka injection leads to activation of mTOR signaling pathway. In this study, we studied the contradictory effect of rapamycin in both normal rats and Ka epileptic rats in more detail. Methods: rats in normal group or Ka model group were given rapamycin at different time points in advance, and brain tissue proteins were extracted at different time points of rapamycin or epileptic seizure at different time points. The protein phosphorylation of Akt, mTOR, Rictor, Raptor, S6K and S6 were analyzed. The behavior of epileptic seizures was monitored and graded according to the modified Racine method. Detection of nerve cell death by Fluoro-Jade B staining Results: we found that in normal rats after injection of rapamycin (3h-24h), it showed the expected dose-dependent inhibition of S6 phosphorylation, but the abnormal increase of S6 phosphorylation level was found at 1 h after administration of rapamycin. In Ka model group, pretreatment with rapamycin more than 10 hours before model making could inhibit the activation of mTOR induced by Ka epilepsy, compared with that of 1h-6h before Ka model making, the activation of mTOR induced by Ka epilepsy could be increased unnaturally. Compared with the control group, the rats pretreated with rapamycin at 1 hour before Ka model had more severe seizure, longer duration and more nerve cell death. Pretreatment with rapamycin 10 hours or more before Ka model had no effect on epilepsy and could reduce the death of nerve cells. This anomalous effect of rapamycin on S6 phosphorylation is consistent with the change of upstream mTOR signal and can be reversed by pirifampicin, a Akt inhibitor. Significance: these data suggest that rapamycin regulates the phosphorylation of S 6 protein and its complexity in epileptic seizures. This anomalous effect should be taken into account in the clinical application of rapamycin as a therapeutic drug for epilepsy and other neuromedical departments.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R965

【參考文獻】

相關(guān)期刊論文 前9條

1 呂偉;張超;;mTOR基因的研究進展[J];重慶醫(yī)學(xué);2005年12期

2 國斌;易斌;魯開智;;哺乳動物雷帕霉素靶蛋白的研究進展[J];醫(yī)學(xué)研究生學(xué)報;2010年08期

3 熊偉;程金湘;章翔;;哺乳動物雷帕霉素靶蛋白研究進展[J];中華神經(jīng)外科疾病研究雜志;2011年01期

4 鄒金凱;車寧;孫春華;;mTOR功能及其抑制劑研究進展[J];現(xiàn)代腫瘤醫(yī)學(xué);2008年10期

5 潘智;張令強;蔣繼志;賀福初;;mTOR的研究進展[J];細(xì)胞生物學(xué)雜志;2006年03期

6 隨力;任杰;;mTOR在記憶功能中的作用[J];中國藥理學(xué)通報;2011年06期

7 劉效磊;牛燕媚;傅力;;mTOR/S6K1信號通路研究進展[J];中國運動醫(yī)學(xué)雜志;2010年01期

8 汝文娟;唐少君;鐘翎;;mTOR信號通路及其與阿爾茨海默病的關(guān)系[J];中國比較醫(yī)學(xué)雜志;2010年08期

9 綦松智;吳登俊;張中顯;;mTOR對信號通路調(diào)控的研究進展[J];中國畜牧雜志;2010年01期

，

本文編號：1785713