本文關(guān)鍵詞：中腦導(dǎo)水管周圍灰質(zhì)結(jié)構(gòu)改變與癲癇發(fā)生發(fā)展的交互影響　出處：《安徽醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 腹側(cè)中腦導(dǎo)水管周圍灰質(zhì) 癲癇 vGLUT1 vGAT

【摘要】：目的通過匹羅卡品腹腔注射誘導(dǎo)大鼠癲癇模型,探索腦腹外側(cè)中腦導(dǎo)水管周圍灰質(zhì)(the ventrolateral Periaqueductal Gray,vPAG)結(jié)構(gòu)改變與癲癇發(fā)生發(fā)展的相互影響。方法將90只達(dá)到既定癲癇持續(xù)狀態(tài)(Status Epilepticus,SE)行為學(xué)標(biāo)準(zhǔn)的大鼠隨機(jī)分成6個(gè)亞組:12h、24h、72h、10d、20d和30d,另隨機(jī)選擇15只與試驗(yàn)組生物學(xué)屬性相似的大鼠作為對(duì)照組。每個(gè)試驗(yàn)亞組在相應(yīng)的時(shí)間點(diǎn)灌注取大鼠腦組織實(shí)施免疫雙標(biāo)熒光染色、HE染色、TUNEL凋亡染色和蛋白印跡(western blot,WB),用以分析vPAG區(qū)神經(jīng)元的囊泡型谷氨酸轉(zhuǎn)運(yùn)體-1(Vesicular Glutamate Transporter1,vGLUT1)和囊泡型γ-氨基丁酸轉(zhuǎn)運(yùn)體(Vesicular GABA Transporter,vGAT)表達(dá)水平的變化。另外隨機(jī)選擇13只正常大鼠用于腦立體定向毀損手術(shù),通過視頻監(jiān)控癲癇行為學(xué)以分析vPAG區(qū)神經(jīng)元的毀損對(duì)自發(fā)性癇性發(fā)作(Spontaneous Recurrent Seizures,SRSs)的影響。試驗(yàn)中所有數(shù)據(jù)均以均數(shù)±標(biāo)準(zhǔn)誤(M±SE)表示,單因素方差分析(One-way ANOVA)用于分析組織化學(xué)染色和WB數(shù)據(jù),獨(dú)立樣本t檢驗(yàn)用于分析癲癇發(fā)作行為學(xué)數(shù)據(jù),以P0.05有統(tǒng)計(jì)學(xué)意義,P0.01有顯著統(tǒng)計(jì)學(xué)意義。結(jié)果在達(dá)到SE后的最初72h內(nèi),vPAG區(qū)神經(jīng)元的vGLUT1和vGAT表達(dá)水平以及平均TUNEL陽性細(xì)胞數(shù)均有明顯的增高趨勢(shì),在SRSs慢性期,上述指標(biāo)呈現(xiàn)出一個(gè)逐漸降低的趨勢(shì),然而直到實(shí)驗(yàn)期結(jié)束時(shí)的30d仍未恢復(fù)至對(duì)照組水平。在達(dá)到SE后的72h,vPAG區(qū)的vGLUT1和vGAT的平均熒光密度顯著高于其他亞組(P0.01),試驗(yàn)組中vPAG區(qū)的平均TUNEL陽性細(xì)胞數(shù)較對(duì)照組明顯增多(P0.01)。在SE后的72h時(shí)間點(diǎn),vPAG區(qū)的vGLUT1和vGAT在WB中的標(biāo)準(zhǔn)化比值較對(duì)照組、12h、20d和30d均有有顯著統(tǒng)計(jì)學(xué)差異(P0.01),此外,在SE后的24h,72h和10d時(shí)間點(diǎn),vPAG區(qū)的vGLUT1和vGAT在WB中的標(biāo)準(zhǔn)化比值均較對(duì)照組有顯著統(tǒng)計(jì)學(xué)差異(P0.01)。vPAG區(qū)受損的神經(jīng)元在HE染色中表現(xiàn)為皺縮、深染的橢圓形細(xì)胞,TUNEL陽性細(xì)胞呈現(xiàn)出褐色或深棕色凝縮致密的細(xì)胞核。經(jīng)過2周的癲癇行為學(xué)視頻監(jiān)測(cè),毀損組中的大鼠癇性發(fā)作頻率、持續(xù)時(shí)間及嚴(yán)重程度較對(duì)照組均有顯著性差異(P0.01)。結(jié)論本研究表明癲癇的發(fā)生發(fā)展誘發(fā)了vPAG功能的失調(diào)和結(jié)構(gòu)的改變,而vPAG結(jié)構(gòu)的損壞也促進(jìn)了癲癇的發(fā)生發(fā)展。
[Abstract]:Objective through the rat model of epilepsy induced by pilocarpine, explore the cerebral ventrolateral periaqueductal gray (the ventrolateral Periaqueductal Gray, vPAG) interaction structure changes and the occurrence and development of epilepsy. Methods 90 rats with Status Epilepticus (SE) behavior criteria were randomly divided into 6 subgroups: 12h, 24h, 72h, 10d, 20d and 30d, and 15 randomly selected rats with similar biological attributes were used as control group. Each test group at the corresponding time point perfused rat brain the immune double fluorescence staining, HE staining, TUNEL staining and Western blotting (western, blot, WB) is used to analyze vPAG neurons of vesicular glutamate transporter -1 (Vesicular Glutamate Transporter1, vGLUT1) and vesicular GABA butyrate transporter (Vesicular GABA Transporter, vGAT) expression. In addition, 13 normal rats were randomly selected for stereotactic brain surgery. The effect of neuronal damage in vPAG area on Spontaneous Recurrent Seizures (SRSs) was analyzed by video surveillance of epileptic behavior. All the data are mean standard error (M + SE), single factor analysis of variance (One-way ANOVA) for the analysis of histochemical and WB data, independent samples t test was used for the analysis of seizure behavior data, using P0.05 statistical significance, P0.01 has significant statistical significance. The results in the first 72h reached SE within vPAG neurons and vGLUT1 expression level of vGAT and the average number of TUNEL positive cells was significantly increased, in the chronic phase of SRSs, the index showed a decreasing trend, however, until the end of the experimental period 30d still did not recover to the level of the control group. After reaching SE, the average fluorescence density of 72h and vPAG in vGLUT1 and vGAT was significantly higher than that in other subgroups (P0.01). The number of TUNEL positive cells in vPAG group was significantly higher than that in control group (P0.01). At 72h time point after SE, vPAG and vGAT in the WB region of vGLUT1 in the standard group, 12h, 20d ratio and 30d were statistically significant compared with the control (P0.01), in addition, after SE, 24h, 72h and 10d time points, vPAG and vGAT in the area of the vGLUT1 standard in WB ratio was higher than the control group with significant difference (P0.01). VPAG area of damaged neurons in HE staining showed blebbing and deep staining oval cells, TUNEL positive cells showed brown or dark brown compact condensed nuclei. After 2 weeks of epileptic behavior video surveillance, the frequency, duration and severity of seizures in the lesioned group were significantly different from those in the control group (P0.01). Conclusion this study showed that the occurrence and development of epilepsy induced the dysfunction of vPAG and the change of structure, and the damage of vPAG structure also promoted the development of epilepsy.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R742.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 孟曙慶;張洪;;難治性癲癇的治療進(jìn)展[J];中華臨床醫(yī)師雜志(電子版);2013年15期

，

本文編號(hào)：1340596