發(fā)布時間：2019-05-28 17:52

【摘要】： 研究目的: 研究重組人粒細胞集落刺激因子(recombinant human granulocyte colonystimulating factor,rhG-CSF)對快速老化模型小鼠P10(senescence acceleratedmouse/prone 10,SAMP10)認知障礙的改善、腦組織炎癥反應的抑制及其抗凋亡作用,為G-CSF治療阿爾茨海默病(Alzheimer disease,AD)的進一步研究提供依據(jù)。 研究方法: 1.隨機將48只SAMP10分為兩組,每組24只:rhG-CSF治療組:rhG-CSF100μg/(kg·d),皮下注射;對照組:等體積生理鹽水,皮下注射。給藥時間均為7d。給藥結束后第7天,14天,28天,56天時間點進行各種指標檢測。 2.認知功能測驗:各組小鼠于給藥前5天及處死前5天進行Morris水迷宮訓練,實驗歷時5天,觀察并記錄其逃避潛伏期和目的象限游泳距離百分比。 3.Morris水迷宮訓練結束后心臟灌注處死小鼠,取腦,制作石蠟切片,進行免疫熒光染色,檢測小鼠額葉腦組織腫瘤壞死因子-α(tumor necrosis factor-α,TNF-α)、誘導型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和半胱氨酸蛋白酶-3(caspase-3)的表達,采用Image-pro Plus圖像分析軟件,分析TNF-α、iNOS和caspase-3陽性細胞百分率、總面積及灰度值。 4.采用SPSS13.0 for Windows軟件包進行統(tǒng)計學處理,各組數(shù)據(jù)以(?)±s表示,差異顯著性檢驗用方差分析和兩樣本t檢驗,P＜0.05為差異有統(tǒng)計學意義。 結果: 1.認知功能測驗 1.1定位航行實驗結果對照組于第14天,28天,56天,逃避潛伏期較給藥前明顯延長(P＜0.05);rhG-CSF治療組于不同時間點與給藥前相比,差異無統(tǒng)計學意義(P＞0.05),第14天逃避潛伏期較對照組縮短(P＜0.05)。 1.2空間探索實驗結果對照組小鼠目的象限游泳距離百分比于給藥后第28天,56天較給藥前減少(P＜0.05);rhG-CSF治療組目的象限游泳距離百分比于各個時間點之間差異無統(tǒng)計學意義(P＞0.05),在第14天,28天較對照組增加(P＜0.05)。 2.免疫熒光染色法檢測額葉TNF-α表達陽性反應物主要分布于大腦額葉皮質,為紅色熒光標記,位于胞漿,細胞核不著色。細胞核用DAPI復染,為藍色熒光標記。對照組治療組于各時間點均可見額葉皮質TNF-α免疫陽性細胞表達,rhG-CSF治療組于第14天、28天、56天TNF-α陽性細胞率,陽性細胞總面積及灰度值均明顯小于對照組(P＜0.05)。 3.免疫熒光染色法檢測額葉iNOS的表達陽性反應物主要分布于大腦額葉皮質,為紅色熒光標記,位于胞漿,細胞核不著色。細胞核用DAPI復染,為藍色熒光標記。對照組治療組于各時間點均可見額葉皮質iNOS免疫陽性細胞表達,iNOS陽性細胞率于治療第14天、28天、56天明顯少于對照組(P＜0.05),陽性細胞總面積于各時間點均小于對照組(P＜0.05)而灰度值于第14天與對照組差異無統(tǒng)計學意義(P＞0.05)。 4.免疫熒光染色法檢測額葉caspase-3表達陽性反應物主要分布于大腦額葉皮質,為紅色熒光標記,位于胞漿亦可見于胞膜胞核。細胞核用DAPI復染,為藍色熒光標記。對照組治療組于各時間點均可見額葉caspase-3免疫陽性細胞表達,陽性細胞率于治療第14天、28天明顯少于對照組(P＜0.05),陽性細胞總面積于28天小于對照組(P＜0.05),灰度值于第28天與對照組有差異(P＜0.05)。 結論: 1.rhG-CSF能夠改善SAMP10的認知障礙,延緩其學習記憶功能的衰退。 2.rhG-CSF能減少SAMP10額葉皮質TNF-α和iNOS陽性細胞表達,抑制小鼠腦組織炎癥反應。 3.rhG-CSF可以減少SAMP10額葉皮質caspase-3陽性細胞的表達,具有抑制神經(jīng)元凋亡的作用。
[Abstract]:The purpose of the study: The effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on the cognitive disorder of mouse P10 (SMP10) in the rapid aging model, the inhibition of inflammatory response of brain tissue and the anti-apoptotic effect of rhG-CSF were studied, and the G-CSF was used to treat Alzheimer's disease (Alzheimer's disease). se, AD) On the basis of. Methods:1.48 SAMP10 were randomly divided into two groups: rhG-CSF (rhG-CSF100. mu.g/ (kg 路 d), subcutaneous injection, control group, etc. Brine water, subcutaneous injection. The administration time was 7 d. Day 7,14,28, and 56 days after the end of the administration 2. Cognitive function test: The Morris water maze training was performed at 5 days before and 5 days before the administration of each group of mice. The experiment lasted for 5 days to observe and record the escape latent period. Objective To determine the percentage of swimming distance in the target quadrant.3. After the end of the Morris water maze training, the mice were sacrificed, the brain was taken, the paraffin section was made, and the immunofluorescent staining was performed to detect the tumor necrosis factor-1 (TNF-1) and the inducible nitric oxide synthase (iNOS) in the frontal lobe of the mouse. Expression of c-oxide synthase (iNOS) and caspase-3 (caspase-3), and image-pro Plus image analysis software was used to analyze the effects of TNF-1, iNOS and caspase-3. Percentage of sex cells, total area, and gray values.4. SPSS13.0 for Windows The software package was statistically processed, and each group of data was expressed in (?)% s, and the variance significance test was performed with an analysis of variance and two different t-tests Inspection, P <0.05 is the difference Results:1. The results were as follows:1. The results of 1.1.1. The experimental results of the cognitive function test (1.1) were significantly prolonged (P <0.05) on the 14th day, the 28th day and the 56 th day, and the rhG-CSF treatment group was different from those of the control group. The difference was not statistically significant (P> 0.0 5) The escape latency of the 14th day was shorter than that of the control group (P <0.05). The percentage of the swimming distance in the control group of the control group in the experimental group was less than that of the control group (P <0.05) on the 28th day after the administration (P <0.05), and the target quadrant of the rhG-CSF treatment group was swimming. The difference between the distance percentage and the time point was not statistically significant (P> 0.05) In day 14 and 28, the control group increased (P <0.05).2. Immunofluorescence staining method for detecting the expression of TNF-antigen in the frontal lobe. The substance is mainly distributed in the frontal cortex of the brain and is a red fluorescent label. In the control group, the expression of TNF-antigen-positive cells in the frontal cortex was observed at all time points, and the rhG-CSF treatment group was in the 14th day, the 28th day, the 56-day TNF-positive-positive cells. The positive cell rate, the total area of positive cells and the gray level were significantly lower than that in the control group (P <0.05).3. The expression of iNOS in the frontal lobe was detected by immunofluorescence staining. The reactive reactant is mainly distributed in the frontal cortex of the brain and is red In the control group, the expression of iNOS, iNOS and iNOS in the frontal cortex were observed in the control group at all time points. The positive cell rate was less than that of the control group (P <0.05) at the 14th day, the 28th day and the 56 th day. The total area of the positive cells was less than that of the control group (P <0.05). The difference of the gray value on the 14th day and the control group was not statistically significant (P> 0.05).4. The caspase-3 in the frontal lobe was detected by the immunofluorescence staining method. The positive reactant is mainly distributed in the frontal cortex of the brain. In the control group, the expression of caspase-3-positive cells in the frontal lobe was observed in the treatment group of the control group. The positive rate of the positive cells in the treatment group was 14 days and 28 days. In less than control group (P <0.05), the total area of positive cells was 28 Day is less than In the control group (P <0.05), the gray value of the control group was different from the control group (P <0.05). Conclusion:1. rhG-CSF can improve the cognitive function of SAMP10 and delay the function of learning and memory. HG-CSF can reduce the expression of TNF-1 and iNOS positive cells in the frontal cortex of the SAMP10, and inhibit the inflammatory reaction of the brain tissue of the mouse.
【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R-332

【相似文獻】

相關期刊論文 前10條

1 ;世界新藥之窗[J];中國新藥雜志;2011年12期

2 ;阿爾茨海默病診斷標準更新[J];浙江中西醫(yī)結合雜志;2011年07期

3 竇麗波;;阿爾茨海默病的住院護理[J];吉林醫(yī)學;2011年26期

4 盛樹力;;表觀遺傳學與阿爾茨海默病研究趨勢[J];中國神經(jīng)免疫學和神經(jīng)病學雜志;2010年06期

5 劉建平;郭蕾;;阿爾茨海默病證候要素研究[J];中西醫(yī)結合心腦血管病雜志;2011年07期

6 張君嗣;楊卓;;阿爾茨海默病Aβ大鼠模型頸上交感神經(jīng)節(jié)神經(jīng)傳導和突觸可塑性的損傷[J];天津醫(yī)藥;2011年07期

7 蓋爾;莊稼英;;糖尿病和腦病[J];糖尿病天地(臨床);2011年06期

8 董浩然;;降壓藥有助于避免老年性癡呆癥[J];心血管病防治知識;2007年12期

9 黃延紅;王俊杰;;阿爾茨海默病的發(fā)病機制及治療[J];濟寧醫(yī)學院學報;2011年03期

10 彭素英;許之駒;李素君;吳燕明;;阿爾茨海默病30例護理效果分析[J];中國民康醫(yī)學;2011年12期

相關會議論文 前10條

1 陳霞平;涂自良;杜士明;王啟斌;張澈;張蓬華;;阿爾茨海默病的藥物治療進展[A];湖北省藥學會第十一屆會員代表大會暨2007年學術年會論文匯編[C];2007年

2 滿永;夏永靜;王抒;;載脂蛋白A-Ⅳ基因360位點與阿爾茨海默病和帕金森病的關系[A];中國病理生理學會動脈粥樣硬化專業(yè)委員會五屆一次會議論文集[C];2002年

3 王乃東;趙永波;;阿爾茨海默病的免疫治療[A];中華醫(yī)學會第七次全國神經(jīng)病學學術會議論文匯編[C];2004年

4 施海姍;侯樂;寧玉萍;鄭東;鐘笑梅;;不同認知障礙的阿爾茨海默病患者睡眠特點分析[A];中華醫(yī)學會精神病學分會第九次全國學術會議論文集[C];2011年

5 金元;曹紅;葛仁山;;Nestorone和黃體酮對阿爾茨海默病大鼠記憶的影響及機制[A];2009年浙江省麻醉學學術會議論文匯編[C];2009年

6 龐麗;姚建莉;李海紅;曹震;徐志峰;尤克增;陳耀文;吳仁華;;9.4T離體型核磁共振頻譜對阿爾茨海默病海馬谷氨酸變化的研究[A];中華醫(yī)學會第16次全國放射學學術大會論文匯編[C];2009年

7 潘小玲;王剛;崔佩菁;王瑛;馬建芳;任汝靜;鄧鈺蕾;徐瑋;湯薈冬;陳生弟;;GAB2基因多態(tài)性與阿爾茨海默病的相關性研究[A];中華醫(yī)學會第十三次全國神經(jīng)病學學術會議論文匯編[C];2010年

8 叢琳;;調控ADAM9基因表達的啟動子區(qū)多態(tài)性對散發(fā)性阿爾茨海默病的保護作用[A];2011全國老年癡呆與衰老相關疾病學術會議第三屆山東省神經(jīng)內(nèi)科醫(yī)師（學術）論壇論文匯編[C];2011年

9 郭U，

本文編號：2487239