expressionsofcd14onperipheralbloodmononuclearcellsfrompatientswithalzheimer39;sdisease

1、中文摘要 中文摘要 研究背景 研究背景 阿爾茨海默病 (Alzheimer’s disease, AD)是一種發(fā)生于中樞神經(jīng)系統(tǒng)的退行性病變。該病的病理學(xué)特征主要涉及Aβ沉積、 Tau 蛋白過(guò)度磷酸化、 炎癥反應(yīng)、 神經(jīng)細(xì)胞凋亡、突觸丟失和顆?？张葑冃裕趸瘧?yīng)激、神經(jīng)細(xì)胞鈣穩(wěn)態(tài)失調(diào)、AD 相關(guān)基因突變及多態(tài)性等機(jī)制相關(guān)。AD原因錯(cuò)綜復(fù)雜，目前，越來(lái)越多的研究證實(shí)，神經(jīng)炎癥機(jī)制在阿爾茨海默病的發(fā)生和發(fā)展進(jìn)程中發(fā)揮了重要作用。在炎性反應(yīng)中，

2、 LPS受體(CD14)與阿爾茨海默病的發(fā)生和發(fā)展進(jìn)程有關(guān)，CD14與LPS結(jié)合， 介導(dǎo)細(xì)胞對(duì)LPS-MD-2與TLR（Toll-like recepter， TLR)4復(fù)合物的識(shí)別,而且， CD14參與小膠質(zhì)細(xì)胞介導(dǎo)的炎性反應(yīng)，調(diào)節(jié)阿爾茨海默病的Aβ攝取和沉積，.我們以前的研究已經(jīng)證實(shí)CD14基因改變AD的遺傳易感性。 因此我們首次提出CD14基因遺傳多態(tài)性可能為AD的遺傳風(fēng)險(xiǎn)因子，而外周血單核細(xì)胞CD14基因和蛋白表達(dá)量可能為AD的

3、早期診斷生物標(biāo)志。 方法 方法 本研究采用美國(guó)神經(jīng)病學(xué)、語(yǔ)言障礙和卒中研究所-老年性癡呆和相關(guān)疾病學(xué)會(huì)(NINCDS—ADRDA)中“很可能 AD(probable)“標(biāo)準(zhǔn)， 選擇 AD 患者 63 例 （年齡≥65 歲，民族為漢族，女性 31 例，平均年齡 74.25 ±15.71 歲），并選擇同期就診的 64 例健康查體者作為對(duì)照（女性 32 例，平均年齡 73.62±15.27 歲）。所有研究對(duì)象均為中國(guó)北方

4、漢族人且無(wú)親緣關(guān)系。 所有入選病人均抽取清晨空腹血各 5.5mLEDTA 抗凝， 應(yīng)用 RNA提取純化試劑盒(美國(guó) Promega)，從外周血單核細(xì)胞提取總 RNA,采用實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)（RT-PCR）技術(shù)、流式細(xì)胞術(shù)分別對(duì) mRNA、血漿蛋白表達(dá)進(jìn)行檢測(cè)?；蚪M DNA 提取按照外周血白細(xì)胞 DNA 提取標(biāo)準(zhǔn)程序,應(yīng)用聚合酶鏈反應(yīng)-限制性片段長(zhǎng)度多態(tài)性(PCR-RFLP)技術(shù),檢測(cè)中國(guó)漢族 AD 人群和健康對(duì)照人群 CD14

5、rs2569190(-159 t / C)和 CD14 rs 2569191(-1145 g / A)基因遺傳多態(tài)性，使用 SPSS 17.0 統(tǒng)計(jì)軟件進(jìn)行統(tǒng)計(jì)分析。AD 組和對(duì)照組各種特征的比較采用 t 檢驗(yàn)或卡方檢驗(yàn)。多組之間的比較采用單因素方差分析，組間兩兩比較采用 Bonferroni 法 ，P<0.05 為差異有統(tǒng)計(jì)學(xué)意義。 所有統(tǒng)計(jì)學(xué)檢驗(yàn)均為雙側(cè)概率檢驗(yàn)， 以 P﹤0.05 為差異有統(tǒng)計(jì)學(xué)意義的標(biāo)Expressions

6、 of CD14 on peripheral blood mononuclear cells from patients with Alzheimer's disease ABSTRACT Background: Alzheimer's disease (AD) is a progressive neurodegenerative disorder，Histopathologically, AD is chara

7、cterized by extracellular amyloid - β (A β ) plaques, intraneuronal neurofibrillary tangles, synaptic loss, neuronal death and microglial activation and recruitment, but its underlying pathological

8、bases are still poorly understood. There is a consensus that inflammatory process has a fundamental role in the pathogenesis of AD. The causes of AD are very complex Currently, there has been increasing evidence

9、suggesting that inflammatory mechanisms are powerful pathogenetic forces in the disease process the innate immune receptors, the LPS receptor (CD14) have been related to AD. The CD14 is pivotal in the initial bindin

10、g of LPS and transfer of LPS to MD2/TLR4 complex to initiate signal cascades Moreover, CD14 are involved in the microglia-mediated inflammatory response, Aβ plaque formation and Aβ clearance in Alzheimer’s disease (AD).

11、Our previous studies have shown that variants in the CD14 genes are associated with the risk of AD. Therefore, we hypothesize that there may be significant changes in CD14 expressions on peripheral blood mononuclear cel

12、ls (PBMCs) from patients with AD when compared to healthy control subjects. Methods: A total of 63 sporadic AD (age at onset ≥ 65 years) patients (women 31; mean age = 73. 38 ±16. 69 years) and 64 healthy controls