本文選題：癲癇　切入點：軸突損害　出處：《重慶醫(yī)科大學(xué)》2017年博士論文　論文類型：學(xué)位論文

【摘要】：第一部分癲癇患者及PTZ點燃癲癇大鼠模型腦組織中的軸突損害目的:探討癲癇患者腦組織和戊四氮(PTZ)點燃癲癇大鼠模型腦組織中軸突損害的情況。方法:收集難治性癲癇患者手術(shù)腦組織標(biāo)本,建立PTZ點燃癲癇大鼠模型并分離收集其腦白質(zhì),用Western blot檢測難治性癲癇患者腦組織和PTZ腹腔注射1天、1周、2周、3周和4周的大鼠腦白質(zhì)中高分子量神經(jīng)細(xì)絲(NF-H)、淀粉樣前體蛋白(APP)、總Tau蛋白、磷酸化Tau蛋白S396(p-Tau S396)和磷酸化Tau蛋白T231(p-Tau T231),并計算Tau蛋白磷酸化水平。結(jié)果:相比對照組,難治性癲癇患者腦組織中NF-H水平明顯降低,APP水平升高,總Tau蛋白減少,Tau蛋白磷酸化水平也升高。PTZ點燃癲癇大鼠腦白質(zhì)中也存在NF-H減少、APP水平升高、總Tau蛋白減少和Tau蛋白磷酸化水平升高,并且其程度隨PTZ注射時間延長而逐漸加重。結(jié)論:難治性癲癇患者和PTZ點燃癲癇大鼠腦組織中均存在以NF-H減少、APP積聚和總Tau蛋白減少為表現(xiàn)的軸突損害,并同時伴有Tau蛋白磷酸化水平升高。第二部分NMDA受體、GSK-3β和Cdk5在癲癇軸突損害中的作用目的:探索NMDA受體、GSK-3β和Cdk5是否參與了癲癇中軸突損害的形成。方法:對大鼠每日腹腔注射PTZ共21天并成功建立點燃癲癇模型后,給予每日腹腔注射非選擇性NMDA受體拮抗劑美金剛、NR2B亞單位選擇性NMDA受體拮抗劑艾芬地爾、GSK-3β拮抗劑氯化鋰、Cdk5拮抗劑Roscovitine或生理鹽水共7天,并用免疫組織化學(xué)術(shù)檢測大鼠胼胝體中NF-H和APP水平,用western blot檢測大鼠腦白質(zhì)中NF-H、APP、Tau、p-Tau S396、和p-Tau T231的水平,并計算Tau蛋白磷酸化水平。結(jié)果:使用美金剛和艾芬地爾阻滯NDMA受體可以減輕PTZ點燃大鼠腦白質(zhì)中NF-H減少和APP積聚的程度,并減輕PTZ點燃導(dǎo)致的總Tau蛋白減少和Tau蛋白磷酸化水平升高。用氯化鋰和Roscovitine分別抑制GSK-3β和Cdk5也可以減輕PTZ點燃導(dǎo)致的腦白質(zhì)中NF-H減少和APP積聚,并減輕PTZ點燃導(dǎo)致的Tau蛋白磷酸化水平升高。此外,氯化鋰可逆轉(zhuǎn)PTZ點燃導(dǎo)致的總Tau蛋白減少,而Roscovitine則并不影響總Tau蛋白的水平。結(jié)論:抑制NMDA受體、GSK-3β和Cdk5均可以改善PTZ點燃癲癇大鼠模型中的軸突損害。提示NMDA受體、GSK-3β和Cdk5興奮參與了癲癇軸突損害的形成。第三部分NMDA受體參與癲癇軸突損害的分子機制目的:研究NMDA受體是否可能通過興奮GSK-3β和Cdk5進而介導(dǎo)癲癇的軸突損害。方法:對大鼠每日腹腔注射PTZ 21天并成功建立點燃癲癇模型后,再行每日腹腔注射非選擇性NMDA受體拮抗劑美金剛、NR2B亞單位選擇性NMDA受體拮抗劑艾芬地爾、GSK-3β拮抗劑氯化鋰、Cdk5拮抗劑Roscovitine或生理鹽水共7天,用western blot檢測大鼠腦白質(zhì)中總GSK-3β、磷酸化GSK-3βY216(p-GSK-3βY216)、磷酸化GSK-3βS9(p-GSK-3βS9)和Cdk5的蛋白水平,用多聚酶鏈?zhǔn)椒磻?yīng)(PCR)檢測大鼠腦白質(zhì)中GSK-3βm RNA和Cdk5 m RNA的水平。結(jié)果:在PTZ點燃大鼠中使用美金剛和艾芬地爾阻滯NDMA受體可以降低Cdk5的m RNA表達和蛋白水平,并且可以升高p-GSK-3βS9、降低p-GSK-3βY216從而抑制GSK-3β的活性,但與美金剛不同的是,艾芬地爾并不影響GSK-3β的m RNA表達和蛋白水平。結(jié)論:NMDA受體,尤其是含NR2B亞單位的NMDA受體,可通過興奮Cdk5和GSK-3β介導(dǎo)癲癇的軸突損害。
[Abstract]:The first part of epilepsy patients and PTZ light axon damage of brain tissues in a rat model of epilepsy Objective: To investigate the brain tissue of patients with epilepsy and four with nitrogen (PTZ) light axonal loss in brain tissue in a rat model of epilepsy. Methods: collecting the brain tissue of patients with refractory epilepsy surgical specimens, the establishment of PTZ kindling rat model of epilepsy the collection and separation of the white matter of the brain, using Western blot to detect brain tissue of patients with refractory epilepsy and PTZ intraperitoneal injection for 1 days, 1 weeks, 2 weeks, the rats of cerebral white matter in high molecular neurofilament 3 and 4 weeks (NF-H), amyloid precursor protein (APP), total Tau protein phosphorylation of Tau protein, S396 (p-Tau S396) and phosphorylated Tau protein T231 (p-Tau T231), and calculate the phosphorylation of Tau protein. Results: compared with control group, NF-H level of brain tissue of patients with refractory epilepsy were significantly decreased, APP increased the level of total Tau protein decreased, Tau protein phosphorylation level also increased.P TZ NF-H also reduced light epileptic rat brain white matter, elevated levels of APP and elevated total Tau protein decreased and Tau protein phosphorylation, and the degree with PTZ injection time gradually increased. Conclusion: the patients with intractable epilepsy and PTZ kindled seizures in the rat brain have to reduce NF-H reduce the accumulation of APP, Tau and total protein of axonal damage, and accompanied by phosphorylation of Tau protein increased. The second part of the NMDA receptor, GSK-3 beta and Cdk5 in axonal damage in epilepsy Objective: To explore the NMDA receptor, GSK-3 beta and Cdk5 is involved in axonal damage in epilepsy. Methods: rats were intraperitoneally injected with PTZ for 21 days and successfully establish a kindling model of epilepsy after given daily intraperitoneal injection of non selective NMDA receptor antagonist memantine, NR2B subunit of NMDA receptor selective antagonist ifenprodil, GSK-3 beta antagonists of lithium chloride, Cdk5 Antagonist Roscovitine or saline for 7 days, and the level of NF-H and APP in rat corpus callosum were detected with immunohistochemistry, Western blot detection of rat brain white matter in NF-H, APP, Tau, p-Tau, S396, p-Tau and T231 levels, and calculate the phosphorylation of Tau protein. Results: the use of memantine and ifenprodil blocking NDMA receptor can reduce PTZ light NF-H cerebral white matter in rats and reduce the degree of accumulation of APP, and reduce PTZ lit caused total Tau protein decreased and Tau protein phosphorylation levels. Brain white matter with lithium chloride and Roscovitine inhibition of GSK-3 beta and Cdk5 can also reduce the PTZ light LED NF-H decrease and APP accumulation, and reduce the level of Tau phosphorylation of PTZ light induced increase. In addition, lithium chloride can lead to the reversal of PTZ kindling total Tau protein decreased, while Roscovitine does not affect the level of total Tau protein. Conclusion: inhibition of NMDA receptor, GSK-3 beta and Cdk5 can improve PTZ kindling axonal injury in rat models of epilepsy. The results indicate that NMDA receptor, GSK-3 beta and Cdk5 in epilepsy excited axonal injury. The purpose of the third part of the NMDA receptor is involved in the molecular mechanism of epilepsy axonal injury: NMDA receptor on the possibility by exciting the GSK-3 beta and Cdk5 mediated axonal damage and epilepsy. Methods: the rats were intraperitoneally injected with PTZ 21 days and successfully establish a kindling model of epilepsy, and daily intraperitoneal injection of non selective NMDA receptor antagonist memantine, NR2B subunit selective NMDA receptor antagonist iseefair to Seoul, GSK-3 beta antagonists of lithium chloride, Cdk5 antagonist Roscovitine or saline 7 day, with total GSK-3 Western blot detection of rat brain white matter in beta, the phosphorylation of GSK-3 Y216 beta (p-GSK-3 beta Y216), phosphorylated GSK-3 S9 beta (p-GSK-3 beta S9) protein and Cdk5, polymerase chain reaction (PCR). Measurement of cerebral white matter in rats with GSK-3 m RNA and Cdk5 m beta RNA level. Results: in PTZ light memantine and ifenprodil blocking NDMA receptor using rat can decrease m RNA expression and protein level of Cdk5, and increased p-GSK-3 beta S9, Y216 inhibits GSK-3 reduce p-GSK-3 beta beta activity. But with memantine is different, ifenprodil did not affect m RNA expression and protein level of GSK-3 beta. Conclusion: the NMDA receptor, especially NR2B subunit containing NMDA receptors, can guide axonal loss in epilepsy by exciting Cdk5 and GSK-3 beta.

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R742.9

【相似文獻】

相關(guān)期刊論文 前5條

1 方煌，羅永湘;化學(xué)發(fā)光法檢測神經(jīng)生長因子軸突逆行運輸?shù)膶嶒炑芯縖J];中華顯微外科雜志;1994年03期

2 赫秋月,韓漫夫,饒明俐;丙烯酰胺對變異Ola鼠和正常6J鼠軸突影響的實驗研究[J];白求恩醫(yī)科大學(xué)學(xué)報;2000年06期

3 戴甲培;王燕子;;軸突轉(zhuǎn)運障礙與阿爾茨海默病[J];中南民族大學(xué)學(xué)報(自然科學(xué)版);2014年02期

4 覃華麗,周雪,章為;生長相關(guān)蛋白GAP-43的研究現(xiàn)狀[J];四川解剖學(xué)雜志;2002年02期

5 王東生;孫洪輝;王心蕊;路來金;;生物素葡聚糖胺觀測周圍神經(jīng)局部軸突運輸?shù)膶嶒炑芯縖J];中華實驗外科雜志;2006年02期

相關(guān)博士學(xué)位論文 前4條

1 劉熙;N-甲基-D-天冬氨酸受體在癲癇軸突損害中的作用及其分子機制研究[D];重慶醫(yī)科大學(xué);2017年

2 鄧敏子;RTN3調(diào)節(jié)BACE1軸突轉(zhuǎn)運的機制研究[D];中南大學(xué);2013年

3 郭效東;軸突及軸突終末變性與再生在C57BL小鼠帕金森病模型中的作用研究[D];第四軍醫(yī)大學(xué);2008年

4 王東生;生物素葡聚糖胺觀測周圍神經(jīng)局部軸突運輸?shù)膶嶒炑芯縖D];吉林大學(xué);2004年

相關(guān)碩士學(xué)位論文 前2條

1 姚晨;化合物SMND-309抗腦缺血/再灌注損傷的體外研究[D];陜西師范大學(xué);2011年

2 李美麗;LPS致炎性大鼠腦內(nèi)AD樣軸突病變的實驗研究[D];中南大學(xué);2014年

，

本文編號：1608870