本文選題：顳葉癲癇　切入點(diǎn)：miR-146a　出處：《延邊大學(xué)》2014年博士論文

【摘要】：研究背景：癲癇是一種常見的神經(jīng)失調(diào)癥,并以反復(fù)自發(fā)異常的神經(jīng)元放電為主要特點(diǎn),目前抗癲癇藥物的使用是治療顳葉癲癇最重要的手段,托吡酯(TPM)則是其中最有效的抗癲癇藥物之一,然而TPM治療癲癇的具體機(jī)制尚未明確。MicroRNAs (miRNAs)一種非編碼單鏈小分子RNA,近幾年的研究表明它不僅對神經(jīng)發(fā)育產(chǎn)生影響,還有研究表明它會參與到癲癇的發(fā)生發(fā)展中,但是其具體機(jī)制尚不明確。結(jié)合國內(nèi)外的許多研究并未發(fā)現(xiàn)miRNAs與TPM治療癲癇機(jī)制的相關(guān)論文,這為明確癲癇發(fā)病機(jī)制和TPM治療癲癇的機(jī)制提供了基礎(chǔ)。本實驗旨在探討microRNAs (miRNAs)在顳葉癲癇中的表達(dá)譜改變,并通過鋰-匹羅卡品顳葉癲癇大鼠模型制備,闡述TPM對于癲癇大鼠海馬凋亡的影響和機(jī)制。 材料和方法：將大鼠隨機(jī)分為四組：1)正常對照組,2)顳葉癲癇組,3)TPM低劑量組(40mg/kg),4) TPM高劑量組(80mg/kg)。通過基因芯片分析技術(shù)測定miRNAs在上述四組中的表達(dá)譜改變,HE染色觀察各組大鼠海馬組織與細(xì)胞的形態(tài)學(xué)改變,并利用免疫蛋白印跡,免疫組化以及免疫熒光等方法測定上述各組海馬中caspase-8蛋白的表達(dá)水平。 結(jié)果：與對照組相比,TLE大鼠海馬中檢測到了12個上調(diào)的miRNAs (miR-146a,-183,-204,-210,-339,-34b,351,-429,-455,466b,-503, and-532)和15個下調(diào)的miRNAs(miR-34a,-383,-451,-101a,-488,-499,-300,-551b,-380,-592,-598,-741,-7b,-374, and-9)。在使用TPM治療后,12個上調(diào)的miRNAs發(fā)生了下調(diào),15個下調(diào)的miRNAs則出現(xiàn)了上調(diào)。 miR-146a是上調(diào)的miRNAs中變化最明顯的。這一結(jié)果闡明,TPM能夠通過主要下調(diào) miR-146a的表達(dá)治療癲癇。另外,在TLE大鼠海馬中可以檢測到caspase-8的表達(dá)水平明 顯增加,但是經(jīng)TPM治療后,其表達(dá)水平下降,證實了TPM能夠通過下調(diào)caspase-8的 表達(dá)抑制TLE大鼠海馬細(xì)胞的凋亡,同時還發(fā)現(xiàn),低劑量TPM處理組中大鼠海馬caspase-8 的表達(dá)較高劑量TPM處理組更低,表明低劑量TPM的對大鼠癲癇的療效比高劑量TPM 更好。通過對大鼠海馬組織上述各組的免疫組化、免疫熒光和HE染色結(jié)果分析發(fā)現(xiàn), 癲癇大鼠的海馬中DG區(qū)的損傷最為嚴(yán)重,其次為CA3區(qū),但是經(jīng)過治療后,其死亡的神 經(jīng)元細(xì)胞明顯減少,證實了TPM的療效顯著,且TPM低劑量的效果較TPM高劑量更為 明顯,這與免疫蛋白印跡的結(jié)果一致。結(jié)論：1)TPM可以通過下調(diào)caspase-8的表達(dá)抑制癲癇小鼠海馬細(xì)胞凋亡的發(fā)生； 2)在TLE的發(fā)生發(fā)展過程中存在miRNAs特異表達(dá)譜,其中miR-146a在TPM治療TLE 過程中具有明顯作用,有望成為TLE治療的分子靶點(diǎn)。
[Abstract]:Background: epilepsy is a common neurological disorder characterized by recurrent spontaneous discharges of neurons. At present the use of antiepileptic drugs is the most important method for the treatment of temporal lobe epilepsy.Topiramate (TPM) is one of the most effective antiepileptic drugs. However, the specific mechanism of TPM in the treatment of epilepsy is not clear.Other studies have shown that it may be involved in the development of epilepsy, but its mechanism is unclear.In combination with many studies at home and abroad, the related papers on the mechanism of miRNAs and TPM in the treatment of epilepsy have not been found, which provides a basis for clarifying the pathogenesis of epilepsy and the mechanism of TPM in the treatment of epilepsy.The purpose of this study was to investigate the changes of microRNAs miRNAs) expression profile in temporal lobe epilepsy and to elucidate the effect and mechanism of TPM on hippocampal apoptosis in rats with lithium-pilocarpine temporal lobe epilepsy.Materials and methods: rats were randomly divided into four groups: 1) normal control group (2)) temporal lobe epilepsy group (30 mg 路kg ~ (-1)) low dose group (40 mg / kg ~ (4)) TPM high dose group (n = 80 mg / kg ~ (-1)).The changes of miRNAs expression profile in the above four groups were detected by gene chip analysis technique. The morphological changes of hippocampus and cells in each group were observed by HE staining, and Western blot was used.Immunohistochemical and immunofluorescence methods were used to detect the expression of caspase-8 protein in hippocampus.緇撴灉:涓庡鐓х粍鐩告瘮,TLE澶ч紶嫻烽┈涓嫻嬪埌浜，

本文編號：1715950