本文選題：縫隙連接蛋白 + 癲癇�。� 參考：《吉林大學(xué)》2014年碩士論文

【摘要】：目的： 癲癇是一種由多種病因引起的慢性腦部疾患，以神經(jīng)元過(guò)度同步放電導(dǎo)致反復(fù)、發(fā)作性和短暫性的中樞神經(jīng)系統(tǒng)功能失常為特征�？p隙連接在癲癇發(fā)病過(guò)程中發(fā)揮重要作用。本實(shí)驗(yàn)擬在建立PTZ誘導(dǎo)的癲癇大鼠模型的基礎(chǔ)上，用縫隙連接通道阻斷劑CBX進(jìn)行干預(yù)處理，觀察記錄各組大鼠行為學(xué)、腦電圖變化，并檢測(cè)癇性發(fā)作后不同時(shí)間點(diǎn)Cx36及Caspase-3、Bcl-2、Bax的表達(dá)變化，進(jìn)一步探討縫隙連接蛋白在癲癇發(fā)作中的作用。 方法： 以大鼠腹腔內(nèi)注射戊四氮制備急性癲癇模型為研究對(duì)象，將大鼠隨機(jī)分為空白對(duì)照組、PTZ組、CBX組。采用HE染色、免疫組織化學(xué)染色、Western blot法等測(cè)定Cx36在癇性發(fā)作后不同時(shí)點(diǎn)(2h、4h、8h、12h、24h)的表達(dá)的變化，并用上述方法檢測(cè)致癇24h后Caspase-3、Bcl-2、Bax在三組間表達(dá)的差異以及DAPI染色檢測(cè)細(xì)胞凋亡。 結(jié)果： 1、PTZ組和CBX組大鼠造模成功，腦電圖均顯示異常，出現(xiàn)棘波、多棘、多尖、尖-慢綜合波等癇性電波。 2、免疫組織化學(xué)染色顯示，大鼠皮質(zhì)以及海馬區(qū)均可以見(jiàn)到大量Cx36免疫陽(yáng)性細(xì)胞，免疫反應(yīng)產(chǎn)物主要分布于神經(jīng)元的胞漿內(nèi)。Cx36表達(dá)變化以海馬CA1和CA3區(qū)為明顯，Cx36蛋白表達(dá)在PTZ和CBX組均從致癇后2h開始增多，并持續(xù)至24h。PTZ組和CBX組分別與對(duì)照組同一時(shí)間點(diǎn)比較，海馬CA1、CA3區(qū)Cx36表達(dá)增多具有統(tǒng)計(jì)學(xué)意義，CBX組與PTZ組各時(shí)間比較，無(wú)顯著差異。 3、Western blot檢測(cè)結(jié)果顯示，Cx36蛋白表達(dá)在PTZ和CBX組均從致癇后2h開始增多，并持續(xù)至24h，與免疫組織化學(xué)染色結(jié)果一致。PTZ組和CBX組大鼠在致癇24h后，，Caspase-3，Bax蛋白表達(dá)增高，Bcl-2蛋白表達(dá)降低，CBX組與PTZ組比較，Caspase-3，Bax蛋白表達(dá)明顯減少，Bcl-2蛋白表達(dá)較多。 結(jié)論： 1． PTZ誘導(dǎo)的急性癲癇大鼠模型中，對(duì)照組模型組的Cx36在皮質(zhì)以及海馬區(qū)均大量表達(dá)。模型組的Cx36的表達(dá)強(qiáng)度明顯高于對(duì)照組。PTZ組與CBX組間表達(dá)無(wú)差異。 2． Cx36表達(dá)及時(shí)程變化以海馬CA1和CA3區(qū)為明顯，在PTZ致癇后2h開始增多，并持續(xù)增多至24h。 3． PTZ誘導(dǎo)的急性癲癇大鼠模型出現(xiàn)神經(jīng)元凋亡，CBX阻斷縫隙連接通道的功能，減輕癲癇導(dǎo)致的神經(jīng)元凋亡的出現(xiàn)。
[Abstract]:Objective: Epilepsy is a kind of chronic brain disease caused by many causes. It is characterized by recurrent, paroxysmal and transient central nervous system dysfunction caused by excessive synchronous firing of neurons. Gap junction plays an important role in the pathogenesis of epilepsy. On the basis of establishing epileptic rat model induced by PTZ, we used gap junction channel blocker CBX to intervene, observe and record the behavior and EEG changes of rats in each group. The expression of Cx36 and Caspase-3 / Bcl-2P Bax were detected at different time points after epileptic seizure to further explore the role of gap junction protein in epileptic seizures. Methods: The acute epilepsy model was established by intraperitoneal injection of pentylenetetrazol in rats. The rats were randomly divided into control group (PTZ) group and CBX group. The expression of Cx36 was detected by HE staining and immunohistochemical staining (Western blot). The expression of Caspase-3, Bcl-2Bcl-2Bax and apoptosis were detected by DAPI staining in 24 hours after epileptic seizure. Results: 1the rats in PTZ group and CBX group were successful in modeling, EEG showed abnormal, and epileptic waves such as spike wave, multi-spine wave, multi-tip wave and cusp-slow complex wave appeared. 2Immunohistochemical staining showed that a large number of Cx36 immunoreactive cells could be seen in the cortex and hippocampus of rats. Immunoreactive products mainly distributed in the cytoplasm of neurons. The expression of Cx36 protein in hippocampal CA1 and CA3 was significantly increased in both PTZ and CBX groups from 2 h after epilepsy, and continued to be compared between 24h.PTZ group and CBX group at the same time point as control group. There was no significant difference in the expression of Cx36 between the PTZ group and the hippocampal CA1 + CA3 group. 3Western blot analysis showed that the expression of Cx36 protein increased in both PTZ and CBX groups from 2 h after epilepsy. The results of immunohistochemical staining were consistent with the results of immunohistochemical staining for 24 h. The expression of Caspase-3 and Bax protein in CBX group was increased 24 hours after epilepsy. The expression of Bcl 2 protein in PTZ group was significantly lower than that in PTZ group, and the expression of Bcl 2 protein was significantly decreased in PTZ group. Conclusion: 1. In the PTZ induced acute epilepsy rat model, the Cx36 in the control group was highly expressed in the cortex and hippocampus. The expression of Cx36 in model group was significantly higher than that in control group and CBX group. 2. The changes of Cx36 expression were observed in the CA1 and CA3 regions of hippocampus, and increased at 2 h after PTZ induced epilepsy, and continued to increase to 24 h after PTZ induced epilepsy. 3. PTZ induced acute epileptic rat model showed the function of neuronal apoptosis and blocked gap junction channel, which alleviated the neuronal apoptosis induced by epilepsy.
【學(xué)位授予單位】：吉林大學(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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