本文選題：阿爾茨海默病 + 殼多糖酶-1�。� 參考：《重慶醫(yī)科大學》2017年碩士論文

【摘要】：第一部分殼多糖酶-1在老年APP/PS1轉(zhuǎn)基因鼠中的表達與認知水平相關目的:殼多糖酶-1(chitinase1,CHIT1)在阿爾茨海默病(Alzheimer’s disease,AD)中表達增加,然而CHIT1在AD中的具體作用尚不明確。本實驗檢測了22月齡APP/PS1雙轉(zhuǎn)基因鼠中CHIT1的表達水平,并結合本團隊既往研究結果,探究CHIT1在AD中的意義。方法:實驗組采用22月齡APP/PS1雙轉(zhuǎn)基因鼠,對照組選取與之相對應的同齡野生型小鼠(wild type,WT)。采用Morris水迷宮檢測行為學變化,聯(lián)合采用酶聯(lián)免疫特異性測定法(enzyme-linked immunospecific assay,ELISA)和實時定量反轉(zhuǎn)錄酶-聚合酶鏈鎖反應(quantitative real-time polymerase chain reaction,q RT-PCR)檢測各組小鼠腦組織中CHIT1的表達水平。結果:與對照組相比,Morris水迷宮顯示22月齡APP/PS1雙轉(zhuǎn)基因鼠逃避潛伏期明顯延長,且在目標象限耗時百分比變短,在非目標象限耗時百分比增加。ELISA法和q RT-PCR顯示22月齡APP/PS1鼠CHIT1表達水平較對照WT組明顯增加。結論:隨著22月齡APP/PS1雙轉(zhuǎn)基因鼠空間學習能力和空間記憶能力的下降,CHIT1表達水平升高。結合本團隊前期關于4月齡、12月齡APP/PS1鼠的實驗結果,AD早期(4月齡APP/PS1鼠)認知下降不突出,AD中晚期(12月齡、22月齡APP/PS1鼠)認知障礙明顯,CHIT1在AD模型中隨著認知功能下降而不斷增高,提示CHIT1可作為評估AD疾病進展的標志物。第二部分殼多糖酶-1在阿爾茨海默病非轉(zhuǎn)基因大鼠模型中的作用機制目的:文獻報道殼多糖酶-1的活性在AD患者中明顯增加,然而CHIT1在AD中的作用機制尚不明確。本實驗旨在探究CHIT1對于AD病理改變以及小膠質(zhì)細胞活性的影響。方法:本研究采用D-半乳糖和Al Cl3持續(xù)注入大鼠腹腔誘導的癡呆模型,并予以外源性的CHIT1和CHIT1抑制劑(chitinase-IN-2),分別檢測CHIT1對于炎癥因子(TNFα,IL-1β,Arg-1,MRC1/CD206)和Aβ寡聚體的影響,Morris水迷宮檢測大鼠行為學改變。此外,進一步采用Aβ處理的N9小膠質(zhì)細胞株以驗證CHIT1對于炎癥因子的影響是通過調(diào)節(jié)小膠質(zhì)細胞活性實現(xiàn)的。結果:無論是D-galactose和Al Cl3誘導的AD動物模型還是Aβ處理的N9小膠質(zhì)細胞株均可檢測到CHIT1活性明顯增高。經(jīng)CHIT1處理的AD大鼠模型中,認知得到改善,同時,還可檢測到抗炎因子(Arg-1,MRC1/CD206)表達水平增高、促炎因子(TNFa,IL-1β)表達水平降低。與之相對應地,在加入CHIT1的Aβ誘導的N9小膠質(zhì)細胞株中,小膠質(zhì)細胞M2抗炎狀態(tài)的標志物表達增多,M1促炎狀態(tài)標志物表達水平下降,表明CHIT1可以調(diào)節(jié)小膠質(zhì)細胞活性,使其向M2抗炎狀態(tài)轉(zhuǎn)換。此外,我們還發(fā)現(xiàn),CHIT1能夠減少Aβ寡聚體在AD大鼠腦組織內(nèi)的沉積。結論:CHIT1可以減少Aβ寡聚體沉積、促使小膠質(zhì)細胞向M2抗炎形態(tài)轉(zhuǎn)化,從而在AD中發(fā)揮保護作用。
[Abstract]:Part I the expression of chitosanase 1 in senile APP/PS1 transgenic mice and its cognitive level objective: the expression of chitinase 1 (CHIT1) in Alzheimer's disease (AD) is increased, but the specific role of CHIT1 in AD is not clear. This study examined the expression of CHIT1 in 22-month-old APP/PS1 transgenic mice and explored the significance of CHIT1 in AD. Methods: the 22 month old APP/PS1 double transgenic mice were used in the experimental group, and the corresponding wild type mice of the same age in the control group were selected. The behavioral changes were detected by Morris water maze, and the expression of CHIT1 in brain tissue of mice in each group was detected by enzyme linked immunospecific assay (Elisa) and real-time quantitative reverse transcriptase polymerase chain reaction quantitative real-time polymerase chain reactionQ (RT-PCRQ). Results: compared with the control group, Morris water maze showed that the escape latency of 22-month-old APP/PS1 transgenic mice was significantly prolonged, and the percentage of time spent in the target quadrant was shortened. The percentage of time consuming in non-target quadrant was increased. Elisa and Q RT-PCR showed that the expression of CHIT1 in 22-month-old APP/PS1 mice was significantly higher than that in control WT group. Conclusion: with the decrease of spatial learning ability and spatial memory ability of 22-month-old APP/PS1 transgenic mice, the expression level of CHIT1 increased. According to the results of our team's previous study on 4-month and 12-month-old APP/PS1 mice, cognitive decline was not significant in AD early and 4-month-old APP/PS1 rats.) Cognitive impairment was significantly increased with cognitive function decline in AD model. The results suggest that CHIT1 can be used as a marker to evaluate the progression of AD. The second part of the mechanism of chitosan enzyme-1 in Alzheimer's disease non-transgenic rats objective: it is reported that the activity of chitosan enzyme-1 is significantly increased in AD patients, but the mechanism of CHIT1 in AD is not clear. The aim of this study was to investigate the effects of CHIT1 on AD pathological changes and microglial activity. Methods: the dementia model was induced by continuous injection of D-galactose and Al Cl3 into the abdominal cavity of rats. The effects of CHIT1 on the inflammatory factor TNF- 偽, IL-1 尾, Arg-1MRC 1 / CD206) and A 尾 oligomer were measured by Morris water maze in rats. In addition, A 尾 -treated N9 microglia cell lines were further used to verify the effect of CHIT1 on inflammatory cytokines by regulating microglial activity. Results: both the AD model induced by D-galactose and Al Cl3 and the N9 microglia cell line treated with A 尾 could significantly increase the activity of CHIT1. In AD rat model treated with CHIT1, the cognition was improved, and the expression of anti-inflammatory factor Arg-1 / MRC1 / CD206 was also detected. In contrast, in A 尾 -induced N9 microglia cell line added with CHIT1, the expression of anti-inflammatory markers of M2 in microglia increased and the expression level of M1 pro-inflammatory markers decreased, suggesting that CHIT1 could regulate microglia activity. Make it to M2 anti-inflammatory state transition. In addition, we also found that CHIT1 can reduce the deposition of A 尾 oligomer in brain tissue of AD rats. ConclusionChIT1 can reduce A 尾 -oligomer deposition, promote microglia to M2 anti-inflammatory morphologic transformation, and thus play a protective role in AD.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R749.16;R-332

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