本文選題：氯化鋰-匹羅卡品癲癇動物模型　切入點：顳葉癲癇　出處：《中南大學》2014年碩士論文　論文類型：學位論文

【摘要】：目的：探討顳葉癲癇發(fā)生過程中海馬神經(jīng)元核膜通透性改變狀況。 方法：將6-8周齡健康雄性SD大鼠114只,隨機分為對照組(n=57只)和實驗組(n=57只),每大組再分為SE后2h、7d和60d三個亞組。應用氯化鋰-匹羅卡品腹腔注射的方法制備癲癇模型,分別進行：1.觀察所有致癇大鼠行為學和腦電圖變化；2.用尼氏染色法觀察大鼠海馬的病理學改變(分別從對照組和實驗組隨機取6只)；3.余大鼠采用蔗糖密度梯度離心技術,分離純化大鼠海馬組織神經(jīng)元細胞核,并觀察分離純化的效果；4.對提取的神經(jīng)元細胞核用免疫熒光法,檢測核膜通透性變化、細胞核凋亡情況,并驗證核膜通透性改變發(fā)生部位與核孔的關系。 結(jié)果： 1.氯化鋰-匹羅卡品大鼠癲癇模型：癲癇造模成功率為70.8%,總死亡率為18.6%。 2.致癇大鼠自發(fā)性癇性發(fā)作：致癇大鼠慢性期均發(fā)生了自發(fā)性癇性發(fā)作。大鼠在SE后8-22天(14.9±6.4天)開始出現(xiàn)自發(fā)性癇性發(fā)作。 3.腦電圖變化：對照組以慢波為主,致癇大鼠急性期腦電圖出現(xiàn)多種形式的癇樣放電,典型波形有尖波、單棘波、多相棘波等發(fā)作性節(jié)律波；靜止期大鼠腦電波亦以慢波為主,未見明顯的癇樣波；慢性期大鼠自發(fā)性癇性發(fā)作的腦電圖可見以尖波為主的癇樣波,且波幅較正常組高。 4.SE后海馬CA1區(qū)、CA3區(qū)和門區(qū)病理學改變：模型組大鼠門區(qū)神經(jīng)元顯著減少,CA1、CA3區(qū)錐體細胞排列紊亂及神經(jīng)元丟失；部分模型大鼠齒狀回顆粒細胞減少、細胞排列紊亂,或顆粒細胞層彌散化。 5.蔗糖密度梯度離心結(jié)果：大鼠海馬組織經(jīng)蔗糖0.32M、2.2M、2.4M、2.6M和2.8M濃度梯度離心后,2.6-2.8M蔗糖層所含細胞核主要是神經(jīng)元細胞核,還有極少量星形膠質(zhì)細胞核。 6.大鼠海馬神經(jīng)元細胞核免疫熒光結(jié)果 (1)核膜通透性檢測結(jié)果：從SE后2h、7d和60d組分離到海馬神經(jīng)元細胞核中,均包括數(shù)量不等的淡綠色陽性細胞核(滲漏細胞核),且顯著多于對照組(P0.01)；且滲漏細胞核的數(shù)量以2h和60d組最多,2h組尤甚(P0.01)。 (2)凋亡細胞核檢測結(jié)果：從SE后2h、7d和60d組分離到海馬神經(jīng)元細胞核,均包括數(shù)量不等的凋亡細胞核。與對照組相比,2h和60d組凋亡細胞核最多,7d組較少(P0.01)。 (3)核膜通透性改變發(fā)生部位與核孔的關系：將SE后2h、7d和60d組大鼠海馬組織中分離純化的神經(jīng)元細胞核分兩份：一份未加核孔阻塞劑麥胚凝集素(WGA),可見不同數(shù)量的發(fā)生核膜滲漏的陽性細胞核。另一份預先加入WGA,各組細胞核均未發(fā)現(xiàn)明顯核膜滲漏。 結(jié)論： 1.SE將導致海馬神經(jīng)元核孔損傷,增加其核膜通透性。 2.核膜通透性增加的存活海馬神經(jīng)元可能參與了顳葉癲癇形成。
[Abstract]:Objective: to investigate the changes of nuclear membrane permeability of hippocampal neurons during temporal lobe epilepsy. Methods: 114 healthy male Sprague-Dawley rats aged 6-8 weeks were randomly divided into control group (n = 57) and experimental group (n = 57). Each group was subdivided into three subgroups: 2 h after SE for 7 d and 60 d. The epileptic model was established by intraperitoneal injection of lithium-pilocarpine. Observe the changes of behavior and electroencephalogram in all epileptic rats 2.The pathological changes of hippocampus of rats were observed by Nissl staining method (6 rats were randomly selected from control group and experimental group respectively. The remaining rats were centrifuged by sucrose density gradient centrifugation. Isolation and purification of neuronal nuclei in rat hippocampal tissue and observation of the effect of isolation and purification 4.The changes of nuclear membrane permeability and nuclear apoptosis were detected by immunofluorescence assay. The relationship between the location of nuclear membrane permeability change and the nuclear pore was verified. Results:. 1. The epileptic model of rats with lithium chloride and pilocarpine: the success rate of epileptic model was 70.8 and the total mortality was 18.6. 2. Spontaneous seizures in epileptic rats: spontaneous epileptic seizures occurred in chronic phase of epileptic rats, and spontaneous seizures began to occur in rats 8-22 days after SE (14.9 鹵6.4 days). 3. EEG changes: the control group was dominated by slow waves, and various forms of epileptiform discharges were observed in the electroencephalogram of epileptic rats in the acute phase. The typical waves included sharp wave, single spike wave, polyphase spike wave and so on. There was no obvious epileptiform wave and the amplitude of epileptiform wave was higher than that of normal group. 4. Pathological changes of hippocampal CA1, CA3 and portal area: in model group, the neuronal disarrangement and neuronal loss in the hippocampal CA1 region were significantly decreased, while in some model rats, the number of granular cells in dentate gyrus decreased and the cell arrangement was disordered. Or granular cell layer dispersion. 5. The results of sucrose density gradient centrifugation: after sucrose concentration gradient centrifugation, the nuclei in sucrose layer of rat hippocampus were mainly neuronal nuclei and a few astrocytes. 6. Results of nuclear immunofluorescence of hippocampal neurons in rats. 1) the results of nuclear membrane permeability test: the nuclei of hippocampal neurons were isolated from the groups of 2 h, 7 d and 60 d after SE. The number of leakage nuclei was significantly higher than that of control group (P 0.01), and the number of leakage nuclei was especially high in 2 h and 60 d groups, especially in 2 h and 60 d groups, especially in 2 h and 60 d groups, especially in 2 h and 60 d groups, especially in 2 h and 60 d groups, especially in 2 h and 60 d groups. (2) the results of apoptotic nucleus detection: hippocampal neuronal nuclei were isolated from 7 d and 60 d groups after SE, including different numbers of apoptotic nuclei. Compared with the control group, the number of apoptotic nuclei in the 2 h and 60 d groups was less than that in the 7 d group. (3) the relationship between the location of nuclear membrane permeability change and the nuclear pores: the neurons isolated and purified from hippocampal tissues of rats in 2 h and 60 d groups after SE were divided into two parts: one without wheat germ agglutinin agglutinin (WGAN), with different numbers. The positive nuclei with nuclear membrane leakage and the other with WGA were not found in each group. Conclusion:. 1. SE will cause the injury of hippocampal neurons' nuclear foramen and increase the permeability of nuclear membrane. 2. Surviving hippocampal neurons with increased nuclear membrane permeability may be involved in the formation of temporal lobe epilepsy.
【學位授予單位】：中南大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R742.1

【參考文獻】

相關期刊論文 前1條

1 盛躍穎;陳敏;;免疫磁珠分離技術在病原微生物檢測中的應用[J];中國食品衛(wèi)生雜志;2011年05期

，

本文編號：1601782