本文選題：實(shí)驗(yàn)性過(guò)敏性腦脊髓炎 + 神經(jīng)功能評(píng)分�。� 參考：《東南大學(xué)》2017年博士論文

【摘要】：研究背景:多發(fā)性硬化(multiple sclerosis, MS)是中樞神經(jīng)系統(tǒng)中最常見(jiàn)的脫髓鞘疾病之一,好發(fā)于青壯年人,起病年齡在20～40歲。流行病學(xué)調(diào)查研究發(fā)現(xiàn)高達(dá)43%到70%的患者存在不同程度的認(rèn)知功能障礙,并且患者的認(rèn)知功能障礙隨著時(shí)間越加嚴(yán)重。認(rèn)知功能障礙嚴(yán)重影響了多發(fā)性硬化患者正常的生活和學(xué)習(xí),給家庭和社會(huì)帶來(lái)了沉重的負(fù)擔(dān)。但是,目前對(duì)多發(fā)性硬化認(rèn)知功能障礙的治療手段有限,療效欠佳,是臨床上亟待解決的問(wèn)題。尸檢和影像學(xué)研究都證實(shí)了多發(fā)性硬化患者認(rèn)知功能活動(dòng)相關(guān)腦區(qū)存在明顯的脫髓鞘損傷,并且研究還發(fā)現(xiàn)髓鞘損傷與認(rèn)知功能障礙密切相關(guān)。然而,髓鞘損傷是否是多發(fā)性硬化認(rèn)知功能障礙的治療靶點(diǎn)還不清楚。含亮氨酸重復(fù)序列和免疫球蛋白結(jié)構(gòu)域的Nogo受體的作用蛋白1(LRR and Ig domain containing, Nogo receptor interacting protein 1,Lingo-1)是一種特異性表達(dá)在中樞神經(jīng)系統(tǒng)中具有抑制少突膠質(zhì)細(xì)胞前體細(xì)胞(OPCs)分化成熟以及髓鞘化的蛋白,而Lingo-1的拮抗劑(Lingo-1抗體)具有促進(jìn)髓鞘修復(fù)的作用。那么LINGO-1抗體是否能夠用于治療多發(fā)性硬化患者認(rèn)知功能障礙目前尚不清楚。本研究中,我們檢測(cè)了兩種常見(jiàn)的多發(fā)性硬化脫髓鞘模型小鼠(實(shí)驗(yàn)性過(guò)敏性腦脊髓炎模型小鼠和Cuprizone誘導(dǎo)的脫髓鞘模型小鼠)的行為學(xué),并用western blot和免疫熒光技術(shù)等觀察了髓鞘蛋白和神經(jīng)元軸突結(jié)構(gòu)和功能蛋白表達(dá)水平以探討:1、脫髓鞘模型小鼠是否存在認(rèn)知功能障礙,以及小鼠認(rèn)知功能相關(guān)腦區(qū)是否存在髓鞘損傷;2、LINGO-1抗體能否改善脫髓鞘模型小鼠的認(rèn)知功能障礙及其病理?yè)p傷。最終,希望通過(guò)以上研究為多發(fā)性硬化認(rèn)知功能障礙的治療提供新思路,為L(zhǎng)INGO-1抗體用于治療多發(fā)性硬化認(rèn)知功能障礙提供依據(jù)。第一部分:實(shí)驗(yàn)性過(guò)敏性腦脊髓炎小鼠的改良與評(píng)價(jià)目的:為了構(gòu)建能夠用于認(rèn)知功能行為學(xué)檢測(cè)的實(shí)驗(yàn)性過(guò)敏性腦脊髓炎(EAE)小鼠,對(duì)EAE造模藥物的劑量進(jìn)行了探索,并對(duì)改良的EAE模型進(jìn)行了評(píng)價(jià)。方法:采用不同劑量的髓鞘少突膠質(zhì)細(xì)胞糖蛋白(MOG35-55) (200μg MOG35-55/只、100μgMOG35-55/只、50μgMOG35-55/只、25μgMOG35-55/只)、滅活結(jié)核分枝桿菌(H37RA)(800 μg滅活結(jié)核分枝桿菌/只、250 μg滅活結(jié)核分枝桿菌/只、100 μg滅活結(jié)核分枝桿菌/只)和百日咳毒素(500 ng百日咳毒素/只、200 ng百日咳毒素/只)等藥物進(jìn)行EAE造模。造模后,按照神經(jīng)功能評(píng)分標(biāo)準(zhǔn)進(jìn)行神經(jīng)功能評(píng)分;用開(kāi)場(chǎng)試驗(yàn)檢測(cè)小鼠自發(fā)活動(dòng)能力。結(jié)果:與高劑量藥物EAE造模組小鼠相比,低劑量藥物造模組小鼠(25μgMOG35-55/只,100 μg滅活結(jié)核分枝桿菌/只,200 ng百日咳毒素/只),在病程中,每天的平均神經(jīng)功能評(píng)分都明顯降低。低劑量藥物造模EAE組小鼠與陰性對(duì)照組相比,自發(fā)活動(dòng)速度無(wú)明顯差別[免疫后第16天,8.885 ±0.772 cm/s (EAE組),8.933±0.567 cm/s (陰性對(duì)照組);免疫后第 31 天,11.130±0.630 cm/s (EAE 組),10.670±0.959 cm/s (陰性對(duì)照組);免疫后第55 天,7.686±0.428 cm/s (EAE 組),8.313±0.918 cm/s (陰性對(duì)照組)](P0.05)。結(jié)論:低劑量藥物(25μgMOG35-55/只,100μg滅活結(jié)核分枝桿菌/只,200ng百日咳毒素/只)所造的EAE模型小鼠存在輕度的神經(jīng)功能障礙,但自發(fā)活動(dòng)功能無(wú)明顯損害。該模型可以用于研究多發(fā)性硬化認(rèn)知功能障礙的病理機(jī)制和藥物開(kāi)發(fā)。第三部分:LINGO-1抗體促進(jìn)髓鞘修復(fù)和EAE小鼠空間學(xué)習(xí)記憶功能恢復(fù)目的:探討LINGO-1抗體能否改善實(shí)驗(yàn)性過(guò)敏性腦脊髓炎模型(EAE)小鼠的認(rèn)知功能障礙(空間學(xué)習(xí)記憶障礙)及其病理?yè)p傷。方法:按照低劑量藥物(25μgMOG35-55/只,100μg滅活結(jié)核分枝桿菌/只,200 ng百日咳毒素/只)誘導(dǎo)EAE小鼠模型;免疫后第14天開(kāi)始系統(tǒng)的給予LINGO-1抗體治療,劑量為10 mg/kg,每6天一次腹腔給藥,給藥直至行為學(xué)檢測(cè)結(jié)束;免疫后第50天開(kāi)始觀察陰性對(duì)第二部分:實(shí)驗(yàn)性過(guò)敏性腦脊髓炎小鼠行為學(xué)和病理表現(xiàn)目的:探討實(shí)驗(yàn)性過(guò)敏性腦脊髓炎(EAE)小鼠是否存在認(rèn)知功能障礙(空間學(xué)習(xí)記憶障礙)和焦慮、抑郁樣行為;探討EAE小鼠認(rèn)知功能相關(guān)的腦區(qū)是否存在脫髓鞘損傷。方法:按照低劑量藥物(25μgMOG35-55/只,100μg滅活結(jié)核分枝桿菌/只,200 ng百日咳毒素/只)誘導(dǎo)EAE小鼠模型,分別在免疫后的早期(第13天)和免疫后的晚期(第55天)進(jìn)行行為學(xué)檢測(cè),包括糖水偏愛(ài)實(shí)驗(yàn)、高架十字迷宮實(shí)驗(yàn)、曠野實(shí)驗(yàn)和水迷宮實(shí)驗(yàn)。在實(shí)驗(yàn)中,每只小鼠只完成其中一個(gè)時(shí)間點(diǎn)的所有行為學(xué)檢測(cè)。行為學(xué)檢測(cè)完成之后,提取腦組織,進(jìn)行western blot檢測(cè)。結(jié)果:EAE小鼠在其病程的早期和晚期,曠野實(shí)驗(yàn)、高架十字迷宮實(shí)驗(yàn)和糖水偏愛(ài)實(shí)驗(yàn)中,表現(xiàn)與陰性對(duì)照組相當(dāng)(P0.05)。水迷宮實(shí)驗(yàn)中,EAE小鼠在病程的早期,表現(xiàn)與陰性對(duì)照組小鼠相當(dāng),不存在空間學(xué)習(xí)記憶障礙(P0.05);但是,EAE小鼠在病程的晚期出現(xiàn)明顯的空間學(xué)習(xí)記憶障礙,表現(xiàn)為學(xué)習(xí)期EAE小鼠找到平臺(tái)的時(shí)間長(zhǎng)于對(duì)照組小鼠,尤其是學(xué)習(xí)期第一天(P0.05),檢測(cè)期EAE小鼠目標(biāo)象限運(yùn)動(dòng)路程百分比明顯低于對(duì)照組的小鼠(P0.05)。Western blot檢測(cè)結(jié)果顯示EAE小鼠的海馬旁回和穹窿-海馬傘出現(xiàn)明顯的脫髓鞘損傷,表現(xiàn)為髓鞘堿性蛋白(MBP)含量下降(P0.05)。結(jié)論:EAE小鼠在病程的晚期出現(xiàn)空間學(xué)習(xí)記憶障礙,這種空間學(xué)習(xí)記憶障礙可能與海馬旁回和穹窿-海馬傘髓鞘損傷有關(guān)。照組、EAE模型組及EAE模型LINGO-1抗體治療組小鼠空間學(xué)習(xí)記憶能力和焦慮抑郁樣行為;行為學(xué)檢測(cè)結(jié)束后,提取腦組織,用western blot和免疫熒光等方法檢測(cè)相應(yīng)腦區(qū)的髓鞘相關(guān)蛋白,軸突結(jié)構(gòu)和功能蛋白以及P13K/AKT/m-TOR信號(hào)通路上蛋白的表達(dá)。結(jié)果:LINGO-1抗體干預(yù)后,三組小鼠在糖水偏愛(ài)實(shí)驗(yàn)、曠野實(shí)驗(yàn)及高架十字迷宮實(shí)驗(yàn)中的表現(xiàn)沒(méi)有顯著差異(P0.05)。而Morris水迷宮實(shí)驗(yàn)中,在學(xué)習(xí)期和檢測(cè)期EAE模型組小鼠的表現(xiàn)明顯差于陰性對(duì)照組小鼠,與陰性對(duì)照組小鼠相比,學(xué)習(xí)期EAE小鼠站臺(tái)潛伏期明顯延長(zhǎng)(P0.05),檢測(cè)期站臺(tái)周圍活動(dòng)路程明顯減少(P0.05) ; LINGO-1抗體治療組小鼠在水迷宮實(shí)驗(yàn)的表現(xiàn)優(yōu)于EAE模型組,與EAE未治療組相比,學(xué)習(xí)期站臺(tái)潛伏期明顯縮短(P0.05),檢測(cè)期站臺(tái)周圍活動(dòng)路程有所增加,其活動(dòng)路程與陰性對(duì)照組相當(dāng)(P0.05)。Western blot檢測(cè)發(fā)現(xiàn)EAE小鼠的海馬旁回和穹窿-海馬傘出現(xiàn)明顯的脫髓鞘損傷,表現(xiàn)為髓鞘堿性蛋白(MBP)的含量下降(P0.05),并伴有軸突順向轉(zhuǎn)運(yùn)蛋白驅(qū)動(dòng)蛋白輕鏈蛋白1(KLC1)含量的下降(P0.05)。給予具有重髓鞘功能的LIINGO-1抗體干預(yù)后,EAE小鼠海馬旁回中的MBP和KLC1明顯升高(P0.05),而穹窿-海馬傘的MBP和KLC1無(wú)明顯提高(P0.05)。此外,LINGO-1抗體治療組小鼠海馬旁回PI3K/AKT/m-TOR信號(hào)通路活化,表現(xiàn)為磷酸化的AKT和磷酸化的m-TOR較EAE未治療組表達(dá)增加(P0.05)。結(jié)論:LINGO-1抗體可能通過(guò)修復(fù)髓鞘改善EAE小鼠的空間學(xué)習(xí)記憶障礙;LINGO-1抗體可能是通過(guò)活化PI3K/AKT/m-TOR信號(hào)通路來(lái)介導(dǎo)這一過(guò)程的;LINGO-1抗體治療可能有助于脫髓鞘疾病患者認(rèn)知功能的恢復(fù)。第四部分:LINGO-1抗體能夠改善Cuprizone誘導(dǎo)脫髓鞘模型小鼠的病理?yè)p傷目的:探討Cuprizone誘導(dǎo)脫髓鞘小鼠行為學(xué)改變以及背側(cè)海馬區(qū)的病理?yè)p傷;探討LINGO-1抗體能否改善Cuprizone誘導(dǎo)脫髓鞘小鼠的行為學(xué)改變及其病理?yè)p傷方法:用0.2% (w/w) cuprizone喂養(yǎng)C57BL/6小鼠誘導(dǎo)脫髓鞘模型,每天喂養(yǎng)直至行為學(xué)檢測(cè)結(jié)束;在造模后第三周開(kāi)始進(jìn)行LINGO-1抗體治療,劑量為10mg/kg,每6天一次腹腔給藥,給藥直至行為學(xué)檢測(cè)結(jié)束;造模后第9周開(kāi)始觀察正常對(duì)照組、cuprizone誘導(dǎo)脫髓鞘模型組、cuprizone誘導(dǎo)脫髓鞘模型LINGO-1抗體治療組三組小鼠空間學(xué)習(xí)記憶能力和焦慮抑郁樣行為;行為學(xué)檢測(cè)結(jié)束后,提取腦組織,采用western blot和免疫熒光等方法檢測(cè)背側(cè)海馬區(qū)的髓鞘相關(guān)蛋白、軸漿轉(zhuǎn)運(yùn)相關(guān)蛋白、軸突結(jié)構(gòu)蛋白、突觸相關(guān)蛋白、tau蛋白、和β淀粉樣蛋白等的表達(dá)。結(jié)果:本研究發(fā)現(xiàn)Cuprizone誘導(dǎo)脫髓鞘模型小鼠存在輕度的空間學(xué)習(xí)能力障礙(P0.05),伴有背側(cè)海馬髓鞘堿性蛋白(MBP)含量下降(P0.05)。給予LINGO-1抗體干預(yù)后,Cuprizone誘導(dǎo)脫髓鞘小鼠記憶能力輕度提高,并伴有MBP含量增加(P0.05)。此外,在Cuprizone誘導(dǎo)脫髓鞘小鼠中,海馬區(qū)同時(shí)有軸突功能蛋白順向轉(zhuǎn)運(yùn)蛋白驅(qū)動(dòng)蛋白1 (KLC1)、軸突結(jié)構(gòu)相關(guān)蛋白神經(jīng)絲蛋白輕鏈(NFL)和神經(jīng)絲蛋白重鏈(NF200)含量顯著下降(P0.05),LINGO-1抗體干預(yù)后,這些軸突功能蛋白和結(jié)構(gòu)蛋白的水平得以提高(P0.05)。然而在Cuprizone誘導(dǎo)脫髓鞘模型小鼠的背側(cè)海馬中,不存在β淀粉樣蛋白水平增加和tau蛋白的過(guò)度磷酸化(P0.05)。結(jié)論:Cuprizone誘導(dǎo)脫髓鞘模型小鼠背側(cè)海馬髓鞘損傷可能參與影響其空間學(xué)習(xí)障礙,LINGO-1抗體可能通過(guò)促進(jìn)髓鞘修復(fù)來(lái)改善其空間學(xué)習(xí)障礙。髓鞘損傷可導(dǎo)致神經(jīng)元軸突破壞,但是可能不會(huì)直接導(dǎo)致β淀粉樣蛋白增加和tau蛋白過(guò)度磷酸化。重髓鞘可能是治療多發(fā)性硬化認(rèn)知功能障礙和恢復(fù)神經(jīng)元功能的一種有效方式。
[Abstract]:Background: multiple sclerosis (MS) is one of the most common demyelinating diseases in the central nervous system, which is well distributed in young people with age of 20~40. Epidemiological studies have found that patients with up to 43% to 70% have different degrees of cognitive dysfunction and cognitive dysfunction with time. More serious. Cognitive dysfunction seriously affects the normal life and learning of patients with multiple sclerosis, which brings a heavy burden to the family and society. However, the treatment of multiple sclerosis is limited and the curative effect is not good. It is an urgent problem to be solved in clinical. Autopsy and imaging studies have confirmed the incidence of multiple sclerosis. There is an obvious demyelinating injury in the brain regions associated with cognitive functional activity in sclerosis, and the study has also found that myelin injury is closely related to cognitive dysfunction. However, it is not clear whether the myelin sheath injury is a target for the treatment of multiple sclerosis cognitive dysfunction. The Nogo receptor containing the repeating sequence of leucine and the immunoglobulin domain The protein 1 (LRR and Ig domain containing, Nogo receptor interacting protein 1, Lingo-1) is a specific expression in the central nervous system that inhibits the maturation and myelination of oligodendrocyte precursor cells (OPCs), and Lingo-1 antagonists have the role of promoting the repair of myelin sheath. It is not clear whether O-1 antibodies can be used in the treatment of cognitive dysfunction in patients with multiple sclerosis. In this study, we detected the behavior of two common multiple sclerosis demyelinating mice (experimental allergic encephalomyelitis model mice and Cuprizone induced demyelinating model mice), and used Western blot and immunofluorescence. The level of myelin protein and neuron axon structure and functional protein expression were observed by optical technology. 1, the presence of cognitive dysfunction in demyelinating model mice, and the presence of myelin injury in the brain regions of cognitive function in mice; 2, whether LINGO-1 antibody could improve the cognitive impairment and pathological damage in demyelinating model mice. Finally, we hope to provide new ideas for the treatment of multiple sclerosis cognitive dysfunction through the above research, and provide the basis for the use of LINGO-1 antibodies in the treatment of multiple sclerosis cognitive impairment. Experimental allergic encephalomyelitis (EAE) mice, the dose of EAE model drugs was explored, and the modified EAE model was evaluated. Methods: using different doses of myelin oligodendrocyte glycoprotein (MOG35-55) (200 mu g MOG35-55/, 100 mu gMOG35-55/ only, 50 u gMOG35-55/ only, 25 u gMOG35-55/), to inactivate Mycobacterium tuberculosis (H37). RA) (800 mu g inactivated Mycobacterium tuberculosis / only, 250 mu g inactivated Mycobacterium tuberculosis / only, 100 mu g inactivated Mycobacterium tuberculosis / only) and pertussis toxin (500 ng pertussis toxin / only, 200 ng pertussis toxin / only) to make EAE model. After making the model, the neural function score was evaluated according to the evaluation criteria of nerve function, and the opening test was used to detect the function. Results: compared with the high dose drug EAE model mice, the low dose drug model mice (25 mu gMOG35-55/ only, 100 micron g inactivated Mycobacterium tuberculosis / only 200 ng pertussis toxin / only), the average daily neural function score in the course of the disease was significantly reduced. The low dose drug model group EAE mice and the negative control group There was no significant difference in the speed of spontaneous activity [sixteenth days after immunization, 8.885 + 0.772 cm/s (EAE), 8.933 + 0.567 cm/s (negative control), thirty-first days after immunization, 11.130 + 0.630 cm/s (EAE group), 10.670 + 0.959 cm/s (negative control group); fifty-fifth days after immunization, 7.686 + 0.428 cm/s (EAE group), 8.313 + cm/s (negative control group)] (P0.05) Conclusion: low dose drugs (25 mu gMOG35-55/ only, 100 mu g inactivated Mycobacterium tuberculosis / only, 200ng pertussis toxin / only) have mild neurological dysfunction, but spontaneous activity has no obvious damage. This model can be used to study the pathological mechanism and drug development of multiple sclerosis recognition dysfunction. Third parts. LINGO-1 antibody promotes myelin repair and EAE mice's spatial learning and memory function recovery purpose: To explore whether LINGO-1 antibody can improve cognitive impairment (spatial learning and memory disorder) and pathological damage in experimental allergic encephalomyelitis model (EAE). Methods: to inactivate tuberculosis branches according to low dose of drug (25 u gMOG35-55/, 100 u G) The EAE mouse model was induced by bacilli / only 200 ng pertussis toxin and only fourteenth days after the immunization, the system was given the LINGO-1 antibody treatment, the dose was 10 mg/kg, every 6 days the intraperitoneal administration, the administration until the behavioral test ended, and the fiftieth days after the immunization, the negative to the experimental allergic encephalomyelitis mice behavior and disease. Objective: To investigate whether there is cognitive dysfunction (spatial learning and memory disorder) and anxiety and depressive behavior in experimental allergic encephalomyelitis (EAE) mice; to explore whether there is demyelinating injury in the brain area related to cognitive function of EAE mice. Methods: a low dose drug (25 u gMOG35-55/ only, 100 mu g inactivated Mycobacterium tuberculosis / only, 20 0 ng pertussis toxin / only induced EAE mice model, respectively, at early (thirteenth days) after immunization and late immunization (fifty-fifth days) to conduct behavioral tests, including sugar water preference experiment, elevated cross maze test, wilderness experiment and water maze test. In the experiment, each mouse only completed all the behavioral tests of one of the time points. After the behavior test was completed, the brain tissue was extracted and the Western blot was detected. Results: the EAE mice in the early and late stages of the disease, the open field experiment, the elevated cross maze experiment and the sugar water preference experiment, were similar to those of the negative control group (P0.05). In the water maze experiment, the EAE mice showed the equivalent of the negative control group in the early stage of the disease. There was no spatial learning and memory disorder (P0.05), but EAE mice had obvious spatial learning and memory disorders in the late course of the disease, which showed that the time of finding the platform in the learning period EAE mice was longer than that of the control group, especially the first day of the learning period (P0.05), and the percentage of the target quadrant movement of the EAE mice was significantly lower than that of the control group. The results of rat (P0.05).Western blot detection showed that the parachyhippocampal and fornix fornix parachute in the EAE mice had obvious demyelination damage, showing the decrease of myelin basic protein (MBP) content (P0.05). Conclusion: the EAE mice appeared in the late stage of the course of the spatial learning and memory disorder, which may be associated with the parahippocampal gyrus and the fornix hippocampus. The spatial learning and memory ability and anxiety and depressive behavior of mice in the EAE model group and the EAE model LINGO-1 antibody treatment group, the brain tissue were extracted after the end of the behavioral test, and Western blot and immunofluorescence were used to detect the myelin related egg white, the axon structure, the functional protein and the P13K/AKT/m-TOR in the corresponding brain region. The expression of protein on the signal pathway. Results: there was no significant difference between the three groups in the glucose water preference experiment, the open field experiment and the elevated cross maze test in the three groups (P0.05). In the Morris water maze experiment, the performance of the mice in the learning period and the detection period of the EAE model group was significantly worse than the negative control group, and the negative was negative. Compared with the group of mice, the incubation period of the EAE mice was obviously prolonged (P0.05), and the activity of the platform around the platform was significantly reduced (P0.05), and the mice in the LINGO-1 antibody treatment group were better than the EAE model group in the water maze test. Compared with the EAE group, the latency of the learning platform was significantly shortened (P0.05), and the activity around the platform platform was observed. The road distance increased, and its activity was similar to that of the negative control group (P0.05).Western blot detection found that the parachyhippocampal and fornix fornix parachute in the EAE mice had obvious demyelination damage, which showed a decrease in the content of myelin basic protein (MBP) (P0.05), accompanied by a decrease in the content of light chain protein 1 (KLC1) of the axon CIS transporter. P0.05). The MBP and KLC1 in the parahippocampal gyrus of EAE mice increased significantly (P0.05) in the parahippocampal gyrus (P0.05) in EAE mice, while the MBP and KLC1 in the fornix hippocampus were not significantly increased (P0.05). Furthermore, the PI3K/AKT/m-TOR signal pathway of the parahippocampal gyrus in the LINGO-1 antibody treatment group was activated by phosphorylated AKT and phosphorylation The expression of EAE untreated group increased (P0.05). Conclusion: LINGO-1 antibody may improve the spatial learning and memory impairment of EAE mice by repairing myelin sheath; LINGO-1 antibody may be mediated through the activation of PI3K/AKT/m-TOR signaling pathway; LINGO-1 antibody therapy may help to restore the cognitive function of demyelinating disease patients. The fourth part LINGO-1 antibody can improve the pathological damage of demyelinating model mice induced by Cuprizone: To explore the behavioral changes of demyelinating mice induced by Cuprizone and the pathological damage in the dorsal hippocampus, and to explore whether LINGO-1 antibody can improve the behavior modification and pathological damage of demyelinating mice induced by Cuprizone: 0.2% (w/w) cuprizone C57BL/6 mice were fed with demyelinating model and were fed every day until the end of behavioral test. Third weeks after the model, the LINGO-1 antibody was treated, the dose was 10mg/kg, the dose was given every 6 days, the drug was given to the end of the behavioral test, and the normal control group was observed at ninth weeks after the model, and the demyelinating model group was induced by cuprizone, cuprizone The three groups of mice were induced by demyelinating model LINGO-1 antibody in space learning and memory ability and anxiety and depressive behavior. After the end of the behavioral test, the brain tissue was extracted. Western blot and immunofluorescence were used to detect the myelin related protein in the dorsal hippocampus, axon transport phase protein, axon structure protein, synapse related protein, tau egg. Results: This study found that the Cuprizone induced demyelinating model mice had a mild spatial learning disability (P0.05), accompanied by a decrease in the content of myelin basic protein (MBP) in the dorsal hippocampus (P0.05). After the prognosis of LINGO-1 antibody, the memory ability of demyelinating mice induced by Cuprizone was slightly improved, accompanied by MBP. Content increased (P0.05). In addition, in Cuprizone induced demyelinating mice, the axon functional protein CIS transporter 1 (KLC1) was found in the hippocampus, and the axon structure related protein neurofilament light chain (NFL) and neurofilament heavy chain (NF200) content decreased significantly (P0.05). The prognosis of LINGO-1 antibody stem, these axon functional proteins and junctions The level of protein structure is improved (P0.05). However, there is no increase in beta amyloid protein level and over phosphorylation of tau protein (P0.05) in the dorsal hippocampus of demyelinating model mice induced by Cuprizone. Conclusion: Cuprizone induced demyelinating mouse dorsal hippocampus myelin injury may be involved in the influence of the spatial learning disability, LINGO-1 antibody. It may improve the spatial learning disorder by promoting myelin repair. Myelin injury may lead to neuronal axonal damage, but it may not directly lead to the increase of beta amyloid and tau protein overphosphorylation.
【學(xué)位授予單位】：東南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R744.51;R-332

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1 周文斌;崔玉真;肖波;;多發(fā)性硬化的流行病學(xué)研究(綜述)[J];中國(guó)神經(jīng)免疫學(xué)和神經(jīng)病學(xué)雜志;2005年06期

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本文編號(hào)：1916091