本文關(guān)鍵詞： P2X3R 顳葉癲癇 興奮性 鈣離子　出處：《重慶醫(yī)科大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:探討P2X3R在難治性顳葉癲癇患者顳葉皮質(zhì)和氯化鋰-匹魯卡品誘導(dǎo)的癲癇大鼠模型腦組織中表達(dá)的變化,通過(guò)膜片鉗全細(xì)胞記錄觀察無(wú)鎂腦脊液誘發(fā)S-D鼠海馬CA1區(qū)錐體神經(jīng)元?jiǎng)幼麟娢活l率加快,對(duì)P2X3R進(jìn)行干預(yù)后,觀察錐體神經(jīng)元?jiǎng)幼麟娢活l率的改變并分析其可能的機(jī)制。方法:1.收集15例難治性顳葉癲癇患者病灶組織標(biāo)本和10例腦外傷手術(shù)患者相對(duì)正常的顳葉皮質(zhì)標(biāo)本。2.稱重后腹腔注射氯化鋰-匹魯卡品誘導(dǎo)S-D大鼠產(chǎn)生急性期癲癇持續(xù)狀態(tài),并通過(guò)視頻監(jiān)控選出2個(gè)月內(nèi)產(chǎn)生自發(fā)發(fā)作的S-D大鼠作為慢性癲癇模型,對(duì)照組腹腔注射生理鹽水進(jìn)行對(duì)照。3.運(yùn)用免疫熒光技術(shù)對(duì)P2X3R在顳葉癲癇患者顳葉組織和癲癇大鼠海馬及鄰近皮質(zhì)的表達(dá)進(jìn)行定位,通過(guò)Western-blot技術(shù)和反轉(zhuǎn)錄聚合酶鏈反應(yīng)(RT-PCR)測(cè)定實(shí)驗(yàn)組與對(duì)照組P2X3R蛋白和相應(yīng)m RNA表達(dá)量的變化。4.S-D鼠麻醉后斷頭取腦,運(yùn)用振動(dòng)切片機(jī)切出含有清晰海馬結(jié)構(gòu)的腦片,腦片厚度為350μm,并在26℃下人工腦脊液孵育1小時(shí),之后運(yùn)用膜片鉗全細(xì)胞記錄技術(shù),通過(guò)激活或抑制P2X3R受體,觀察無(wú)鎂狀態(tài)下誘發(fā)的快速動(dòng)作電位頻率的變化。結(jié)果:P2X3R在顳葉癲癇患者顳葉皮質(zhì)和癲癇大鼠海馬及鄰近皮質(zhì)的神經(jīng)元樹(shù)突和胞質(zhì)中均有表達(dá),并且P2X3受體在癲癇患者及癲癇大鼠模型中表達(dá)升高。無(wú)鎂腦脊液可誘發(fā)S-D鼠海馬CA1區(qū)錐體神經(jīng)元興奮性升高,引起快速樣放電,抑制P2X3R受體后,錐體神經(jīng)元?jiǎng)幼麟娢活l率減慢,激活受體后,電活動(dòng)加快。結(jié)論:P2X3R在癲癇患者及癲癇大鼠模型中表達(dá)升高并且阻斷P2X3R可抑制神經(jīng)元過(guò)高的興奮性,故我們推測(cè)升高的P2X3R在癲癇的形成過(guò)程中具有一定作用。
[Abstract]:Objective: to investigate the changes of P2X3R expression in the temporal cortex of patients with refractory temporal lobe epilepsy and the brain tissue of rats with epilepsy induced by lithium-pilocarpine. The frequency of action potential of pyramidal neurons in hippocampal CA1 region of S-D rats induced by magnesium free cerebrospinal fluid (CSF) was observed by whole-cell patch clamp recording. To observe the change of action potential frequency of pyramidal neurons and to analyze its possible mechanism. Methods: 1. 15 patients with intractable temporal lobe epilepsy and 10 patients with traumatic brain surgery were collected. After weighing, S-D rats were induced by intraperitoneal injection of lithium chloride and pilocarpine to produce acute epileptic status. S-D rats with spontaneous seizures within 2 months were selected as chronic epilepsy models by video surveillance. The expression of P2X3R in temporal lobe tissues of temporal lobe epilepsy and hippocampus and adjacent cortex of epileptic rats was determined by immunofluorescence technique. The changes of P2X3R protein and corresponding m RNA expression were measured by Western-blot and reverse transcriptase polymerase chain reaction (RT-PCR). 4. After anesthesia, brain slices with clear hippocampal structure were cut out by vibration slicer. The thickness of brain slices was 350 渭 m, incubated with artificial cerebrospinal fluid at 26 鈩，

本文編號(hào)：1503735