本文選題：miR-132　切入點(diǎn)：antagomir　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p-CREB/miR-132在顳葉癲癇患者及動(dòng)物模型中的表達(dá) 目的：探討p-CREB和miR-132在難治性癲癇患者致癇灶的表達(dá)以及p-CREB和miR-132在氯化鋰-匹羅卡品癲癇模型中海馬組織中各時(shí)間點(diǎn)的表達(dá)規(guī)律。 方法： 1.選擇15例難治性癲癇患者顳葉手術(shù)切除標(biāo)本和性別、年齡無(wú)差異的10例非癲癇患者顳葉組織標(biāo)本。 2.選擇成年SD大鼠誘導(dǎo)氯化鋰-匹羅卡品模型，隨機(jī)分為2組：正常對(duì)照組，癲癇模型組，根據(jù)癲癇進(jìn)程分為7個(gè)亞組（6h，24h，3d，7d，14d，30d，60d）。 3.用westernblot技術(shù)和免疫組化技術(shù)檢測(cè)p-CREB的表達(dá)水平，實(shí)時(shí)熒光定量PCR檢測(cè)miR-132的表達(dá)水平。 結(jié)果： 1.p-CREB在顳葉癲癇患者中及大鼠癲癇模型中各時(shí)間點(diǎn)表達(dá)均明顯升高。 2.miR-132表達(dá)在顳葉癲癇患者組中表達(dá)降低（p＜0.05），在動(dòng)物模型中24h、7d時(shí)間點(diǎn)miR-132表達(dá)顯著升高（p＜0.05），其余時(shí)間點(diǎn)較正常組無(wú)顯著性差異。 結(jié)論：p-CREB/miR-132的表達(dá)癲癇起病的急性期和潛伏期表達(dá)升高，，呈平行關(guān)系，這條信號(hào)通路早期可能參與了癲癇的發(fā)生和發(fā)展過(guò)程。 第二部分抑制miR-132的表達(dá)對(duì)大鼠癲癇發(fā)作的作用及機(jī)制 目的：通過(guò)抑制miR-132的表達(dá)，研究其在癲癇發(fā)病機(jī)制中的相關(guān)作用。 方法： 1.選擇成年SD大鼠，分為兩組Ant-132干預(yù)組和Scr陰性對(duì)照組，造模前預(yù)先立體定位腦室分別注射Ant-132和Scr，濃度分別為0.5nmol、1.0nmol、1.5nmol。2天后進(jìn)行誘導(dǎo)氯化鋰-匹羅卡品顳葉癲癇模型。分析其對(duì)癲癇模型造模過(guò)程的影響。 2.選擇1.0nmol的Ant-132和Scr作為干預(yù)和控制濃度，分別獲取24h、2w、1m組織標(biāo)本。 3.行為學(xué)觀測(cè)干預(yù)miR-132后，對(duì)大鼠癲癇發(fā)作的影響。NPY檢測(cè)對(duì)苔蘚纖維出芽的影響。Golgi染色檢測(cè)在活體組織干預(yù)miR-132后對(duì)神經(jīng)元樹(shù)突形態(tài)的影響。 結(jié)果： 1.在匹羅卡品造模的過(guò)程中，抑制miR-132表達(dá)可以延長(zhǎng)誘導(dǎo)大鼠癲癇發(fā)作的時(shí)間，并與miR-132濃度呈正相關(guān)（p＜0.05）。 2.抑制miR-132可以有效降低慢性期癲癇的自發(fā)性發(fā)作次數(shù)（p＜0.05）。 3.抑制miR-132后，苔蘚纖維出芽顯著減少，逆轉(zhuǎn)性降低CA3、CA1區(qū)樹(shù)突出芽，降低該區(qū)樹(shù)突的長(zhǎng)度。（p＜0.05） 結(jié)論：miR-132在癲癇的發(fā)生發(fā)展過(guò)程中起了重要作用。MiR-132為癲癇發(fā)病過(guò)程中重要的致病因子。抑制miR-132可能通過(guò)重塑MFs-CA3環(huán)路，減少神經(jīng)環(huán)路的產(chǎn)生從而達(dá)到有效減緩癲癇發(fā)生的作用。
[Abstract]:The expression of p-CREBR / miR-132 in patients with temporal lobe epilepsy and animal models. Aim: to investigate the expression of p-CREB and miR-132 in epileptogenic foci in patients with intractable epilepsy and the expression of p-CREB and miR-132 in hippocampus of lithium-pilocarpine epileptic model. Methods:. 1. The temporal lobe tissue specimens of 15 patients with intractable epilepsy and 10 patients with no difference in age were selected. 2. Adult SD rats were selected to induce lithium-pilocarpine model and were randomly divided into two groups: normal control group and epileptic model group. 3. The expression of p-CREB was detected by westernblot and immunohistochemistry, and the expression of miR-132 was detected by real-time fluorescence quantitative PCR. Results:. 1. The expression of p-CREB was significantly increased in temporal lobe epilepsy patients and rat epileptic models at all time points. 2. The expression of miR-132 in the patients with temporal lobe epilepsy decreased significantly (P < 0.05), and the expression of miR-132 increased significantly at 24 h and 7 d in the animal model, but there was no significant difference between the other time points and the normal group. Conclusion the expression of the expression of the fraction p-CREB-miR-132 is increased in the acute phase and latent period of epileptic onset, which may be involved in the occurrence and development of epilepsy in the early stage. The second part: inhibitory effect of miR-132 expression on epileptic seizures in rats and its mechanism. Objective: to study the role of miR-132 in the pathogenesis of epilepsy by inhibiting its expression. Methods:. 1. Adult SD rats were divided into two groups: Ant-132 intervention group and Scr negative control group. The temporal lobe epilepsy model of lithium-pilocarpine was induced by intracerebroventricular injection of Ant-132 and scratch at a concentration of 0.5 nmol ~ 1.0 nmol ~ (-1) nmol ~ (2) 2 days before modeling, and the effects on the process of epileptic model were analyzed. 2.1.0 nmol of Ant-132 and Scr were selected as the intervention and control concentration, and the tissue samples of 24 h and 2 weeks were obtained, respectively. 3.Behavioral observation after intervention in miR-132, the effect of NPY on the sprouting of mossy fibers in rats. Golgi staining was used to detect the effect of miR-132 in vivo on the morphology of neuronal dendrites. Results:. 1. In the course of pilocarpine modeling, inhibiting the expression of miR-132 could prolong the time of epileptic seizure induced by pilocarpine in rats, and had a positive correlation with the concentration of miR-132 (p < 0.05). 2. Inhibition of miR-132 can effectively reduce the number of spontaneous seizures in chronic epilepsy (p < 0.05). 3. After inhibiting miR-132, the sprouting of moss fiber decreased significantly, the reverse effect decreased, and the length of dendrites in CA3 / CA1 region was decreased, and the length of dendrites in this area was decreased (P < 0.05). Conclusion: miR-132 plays an important role in the occurrence and development of epilepsy. MiR-132 is an important pathogenic factor in the pathogenesis of epilepsy. The inhibition of miR-132 may reduce the generation of neural loop by remodeling the MFs-CA3 loop and thus reduce the occurrence of epilepsy effectively.
【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)】：R742.1

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相關(guān)期刊論文 前1條

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