本文選題：癲癇 + 經(jīng)皮三叉神經(jīng)電刺激　； 參考：《安徽醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的： 1.觀察經(jīng)皮三叉神經(jīng)電刺激（TcTNS）對(duì)匹羅卡品誘發(fā)的癲癇持續(xù)狀態(tài)（SE）所致海馬神經(jīng)元損傷的保護(hù)作用。 2.觀察觀察經(jīng)皮三叉神經(jīng)電刺激（TcTNS）對(duì)匹羅卡品誘發(fā)的慢性癲癇模型大鼠海馬神經(jīng)元GAD65/67表達(dá)的影響，以探討經(jīng)皮三叉神經(jīng)電刺激治療癲癇及腦保護(hù)作用的可能機(jī)制。 資料和方法： 實(shí)驗(yàn)一： 選用健康成年雄性SD大鼠，采用隨機(jī)數(shù)字表法將大鼠隨機(jī)分為二組，匹羅卡品模型組和對(duì)照組，模型組用匹羅卡品誘導(dǎo)癲癇點(diǎn)燃模型，對(duì)照組注射等量的生理鹽水，經(jīng)過行為學(xué)觀察兩周后，具有慢性癲癇自發(fā)性再發(fā)作的為模型制備成功大鼠，再將模型制備成功大鼠采用隨機(jī)數(shù)字表法分為兩組，治療組和模型組，分別給予三叉神經(jīng)電刺激和假刺激一個(gè)月（刺激參數(shù)為頻率140Hz,電流10mA,脈寬0.5ms，正向脈沖刺激30s間歇5min，連續(xù)刺激1h，假刺激組各項(xiàng)參數(shù)均為0。刺激時(shí)間定為每天12:00-16:00）后終止刺激，再次注射皮羅卡品誘導(dǎo)一次癲癇持續(xù)狀態(tài)（SE）并于SE后24h,48h,72h處死大鼠并取材，對(duì)照組亦在相應(yīng)時(shí)間點(diǎn)取材，采用TUNNEL和NISSEL染色觀察各組大鼠海馬神經(jīng)元細(xì)胞的凋亡和壞死情況。組間比較采用配對(duì)t檢驗(yàn)，以p㩳0.05定義為差別有統(tǒng)計(jì)學(xué)意義。 實(shí)驗(yàn)二： 選用健康成年雄性SD大鼠，采用同實(shí)驗(yàn)一的方法建立癲癇點(diǎn)燃模型，將慢性癲癇模型制備成功者用隨機(jī)數(shù)字表法分為模型組與治療組，分別給予三叉神經(jīng)電刺激與假刺激一個(gè)月（刺激參數(shù)及時(shí)間同實(shí)驗(yàn)一），于停止刺激后24h，72h，一周，二周，四周處死大鼠并取材，對(duì)照組亦于相應(yīng)時(shí)間點(diǎn)取材。采用免疫組化的方法研究大鼠海馬及皮層GAD65、67的表達(dá)情況，組間比較采用配對(duì)t檢驗(yàn)，組內(nèi)比較采用單因素方差分析，以p㩳0.05定義為差別有統(tǒng)計(jì)學(xué)意義。 結(jié)果： 1．在實(shí)驗(yàn)一中： 1.1TUNNEL染色：SE后各時(shí)間點(diǎn)治療組和模型組均可見TUNNEL陽性細(xì)胞，主要分布在海馬的CA1和CA3區(qū)，SE后24h，48h，72h治療組CA3區(qū)的TUNNEL陽性細(xì)胞較模型組顯著減少（p＜0.05），以72小時(shí)尤為明顯（p＜0.01）正常對(duì)照組可見少量TUNNEL陽性細(xì)胞。 1.2NISSEL染色：正常對(duì)照組顯示海馬CA1和CA3區(qū)神經(jīng)元細(xì)胞排列整齊緊密，沒有明顯的神經(jīng)元缺失，細(xì)胞核呈圓形或橢圓形，胞漿內(nèi)尼氏小體豐富。模型組各時(shí)間點(diǎn)可見不同程度的神經(jīng)元丟失，部分神經(jīng)元胞體皺縮，胞漿深染，核固縮，錐體細(xì)胞存活數(shù)較正常細(xì)胞減少，尤以72小時(shí)最為明顯（p＜0.01）。經(jīng)皮三叉神經(jīng)刺激治療組各時(shí)間點(diǎn)大鼠海馬CA1，，CA3區(qū)神經(jīng)元細(xì)胞尚清晰完整，部分細(xì)胞出現(xiàn)染色質(zhì)凝集，神經(jīng)元丟失情況較模型組顯著下降，即治療組Nissl染色細(xì)胞數(shù)顯著多于模型組（p＜0.05）。 2.在實(shí)驗(yàn)二中： 2.1：海馬區(qū)及皮層均有GAD65的陽性細(xì)胞表達(dá)，尤以海馬的CA3區(qū)表達(dá)最為明顯，主要在胞核中表達(dá)，陽性細(xì)胞為棕紅色顆粒，經(jīng)皮三叉神經(jīng)刺激治療組較假刺激組相比，各時(shí)間點(diǎn)治療組陽性細(xì)胞表達(dá)均高于模型組（p＜0.05），治療組GAD65的表達(dá)于停止刺激后72小時(shí)-7天達(dá)到高峰，較模型組有極顯著差異（p＜0.01）。后GAD65的表達(dá)下降，至28天接近正常表達(dá)水平。 2.2：海馬區(qū)及皮層均可見GAD67陽性細(xì)胞，在海馬的CA3區(qū)及DG區(qū)表達(dá)最為明顯，主要在胞漿中表達(dá)，與模型組各時(shí)間點(diǎn)相比較，治療組的表達(dá)遠(yuǎn)大于模型組（p＜0.05）。組內(nèi)比較發(fā)現(xiàn)，GAD67無明顯的時(shí)間變化趨勢，至停止刺激后28天，治療組的GAD67的表達(dá)仍大于正常水平。 結(jié)論： 1.經(jīng)皮三叉神經(jīng)電刺激治療可以減少癲癇大鼠海馬神經(jīng)元的損傷，從而發(fā)揮對(duì)癲癇大鼠海馬神經(jīng)元的保護(hù)作用。 2.經(jīng)皮三叉神經(jīng)電刺激可使癲癇大鼠腦內(nèi)抑制性神經(jīng)遞質(zhì)的標(biāo)志物GAD65/67表達(dá)上調(diào)，,抑制性遞質(zhì)的增強(qiáng)可能為三叉神經(jīng)電刺激治療癲癇及發(fā)揮腦保護(hù)作用的機(jī)制之一。
[Abstract]:Purpose :

1 . To observe the protective effect of transcutaneous trigeminal nerve stimulation ( TcN ) on hippocampal neuronal injury induced by pilocarpine ( SE ) .

2 . To observe the effect of percutaneous trigeminal nerve stimulation ( TcN ) on the expression of GAD65 / 67 in rat hippocampal neurons induced by pilocarpine , and to investigate the possible mechanism of percutaneous trigeminal nerve stimulation in the treatment of epilepsy and brain protection .

Information and methods :

Experiment 1 :

Adult male SD rats were randomly divided into two groups : two groups , pilocarpine model group and control group . The model group was divided into two groups by pilocarpine . After two weeks of behavioral study , the rats were divided into two groups : treatment group and model group .

Experiment 2 :

The rats were randomly divided into three groups : model group and treatment group . The rats were sacrificed at 24 h , 72 h , one week , two weeks and four weeks after cessation of stimulation . The expression of GAD65 and 67 in hippocampus and cortex of rats was studied by immunohistochemistry .

Results :

1 . In Experiment 1 :

1 . After SE treatment group and model group , positive cells were observed , and the number of positive cells was significantly decreased at 24 h , 48 h and 72 h after SE ( p < 0 . 05 ) , and a small number of positive cells were found in the normal control group at 72 hours ( p < 0.01 ) .

1 . 2NISSEL staining : The normal control group showed that the neuronal cells in the CA1 and CA regions of the hippocampus were orderly and compact , no obvious neuronal loss was found , the nuclei were round or oval , and the Nissl in the cytoplasm was abundant .

2 . In Experiment 2 :

2.1 : The expression of GAD65 positive cells in the hippocampus and cortex was the most obvious , especially in the CA 3 region of hippocampus . The positive cells were higher than those in the model group ( p < 0.05 ) . The expression of GAD65 in the treatment group was higher than that in the model group ( p < 0.05 ) . The expression of GAD65 in the treatment group decreased from 72 hours to 7 days after cessation of stimulation , and the expression of GAD65 decreased to close to normal expression level 28 days .

2.2 : The expression of GAD67 positive cells was found in the hippocampus and in the cortex . The expression of GAD67 in hippocampus was much higher than that in model group ( p < 0.05 ) . Compared with model group ( p < 0.05 ) , the expression of GAD67 was much larger than that in model group ( p < 0.05 ) .

Conclusion :

1 . Percutaneous trigeminal nerve stimulation can reduce the damage of hippocampal neurons of epileptic rats , and play an important role in the protection of hippocampal neurons in epileptic rats .

2 . The expression of GAD65 / 67 is up - regulated by percutaneous trigeminal nerve stimulation , and the enhancement of inhibitory neurotransmitter may be one of the mechanisms of trigeminal nerve stimulation in the treatment of epilepsy and brain protection .
【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】：2014
【分類號(hào)】：R742.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 張慧敏;李良勇;李家林;王玉;;經(jīng)皮三叉神經(jīng)電刺激預(yù)處理對(duì)戊四氮致癇大鼠海馬谷氨酸脫羧酶表達(dá)的影響[J];安徽醫(yī)科大學(xué)學(xué)報(bào);2011年08期

2 王煥明,譚啟富,吳波,印紅霞;顳葉癲癇患者腦內(nèi)細(xì)胞凋亡現(xiàn)象的研究[J];中華神經(jīng)外科雜志;1999年06期

3 張軍強(qiáng);余巨明;王曉明;趙紅寧;黃敏;張小東;趙曉瓊;黃慧;胡建秀;;低頻重復(fù)經(jīng)顱磁刺激預(yù)處理對(duì)毛果蕓香堿致癇大鼠海馬GAD65、NMDAR1表達(dá)的影響[J];中國神經(jīng)免疫學(xué)和神經(jīng)病學(xué)雜志;2008年06期

本文編號(hào)：2013970