本文選題：細(xì)粒棘球蚴 + 熒光成像��； 參考：《石河子大學(xué)》2016年碩士論文

【摘要】：目的:1.嘗試細(xì)粒棘球蚴體外熒光染色,通過紅綠熒光信號(hào)強(qiáng)度分析不同藥物對(duì)原頭蚴活性的影響。2.構(gòu)建細(xì)粒棘球蚴小鼠活體內(nèi)熒光發(fā)光模型,并對(duì)小鼠肝臟中的細(xì)粒棘球蚴感染灶進(jìn)行定位及量化分析。探討構(gòu)建小鼠活體內(nèi)生物發(fā)光模型的可能性。方法:1.無菌分離提取原頭蚴,將原頭蚴分為對(duì)照組、二甲雙胍處理組(將10 m M二甲雙胍加入原頭蚴培養(yǎng)基中)、聯(lián)合用藥處理組(將10 m M二甲雙胍以及15μM阿苯達(dá)唑(相當(dāng)于4.2μg/m L)聯(lián)合加入原頭蚴培養(yǎng)基中)。培養(yǎng)并動(dòng)態(tài)監(jiān)測原頭蚴活性。2.采用JC-1熒光染料對(duì)不同處理組原頭蚴進(jìn)行熒光染色,并置于共聚焦熒光顯微鏡下采集成像并記錄分析各組原頭蚴熒光強(qiáng)度。3.將不同處理組的原頭蚴置于24孔板中,連續(xù)培養(yǎng)并采用小動(dòng)物發(fā)光成像儀對(duì)三組原頭蚴熒光強(qiáng)度進(jìn)行動(dòng)態(tài)監(jiān)測,直至熒光信號(hào)消失。4.將不同處理組的原頭蚴染色后種植于CF-1雄性小鼠肝臟被膜下,將不同注射組的小鼠置于小動(dòng)物發(fā)光成像儀中進(jìn)行動(dòng)態(tài)成像分析。選取對(duì)照組小鼠進(jìn)行熒光梯度實(shí)驗(yàn),分析熒光信號(hào)衰減趨勢。結(jié)果:1.與對(duì)照組相比,Met藥物處理組原頭蚴活性出現(xiàn)一定程度的下降,ABZSO和Met藥物聯(lián)合使用組原頭蚴活性下降最為顯著。2.對(duì)照組原頭蚴的紅色熒光強(qiáng)度明顯強(qiáng)于綠色熒光;Met藥物處理組原頭蚴紅色熒光強(qiáng)度與綠色熒光沒有顯著差異;ABZSO和Met藥物聯(lián)合使用組原頭蚴綠色熒光強(qiáng)度明顯強(qiáng)于紅色熒光。3.三組中原頭蚴的紅色熒光出現(xiàn)了明顯的下降趨勢,聯(lián)合用藥組的下降趨勢最為明顯,其次是二甲雙胍藥物處理組,對(duì)照組的下降趨勢較為平緩。而綠色熒光在聯(lián)合用藥組與二甲雙胍處理組中出現(xiàn)了明顯的增強(qiáng)趨勢,且聯(lián)合用藥組強(qiáng)于二甲雙胍組,隨后相應(yīng)出現(xiàn)下降的趨勢。對(duì)照組的綠色熒光從檢測開始就處于較低水平,且有一定的下降趨勢,實(shí)驗(yàn)組與對(duì)照組相比具有顯著的統(tǒng)計(jì)學(xué)差異(P0.05)。4.成功種植進(jìn)小鼠肝臟后,對(duì)照組的紅色熒光區(qū)域明顯大于綠色熒光,且紅色熒光強(qiáng)度強(qiáng)于綠色熒光,紅/綠熒光的強(qiáng)度比為4.1。二甲雙胍組的紅色熒光信號(hào)與綠色熒光相比,無論是熒光區(qū)域的大小還是熒光強(qiáng)度都沒有明顯的差異,且紅/綠比值為1.2。聯(lián)合用藥組的紅色熒光區(qū)域要小于綠色熒光,且紅色熒光強(qiáng)度也弱于綠色熒光,紅/綠比值為0.5。對(duì)照組小鼠肝區(qū)的紅色熒光在觀察初期強(qiáng)度較高,隨后逐漸的減弱。綠色熒光在種植后出現(xiàn)一定范圍內(nèi)的上升趨勢,隨后逐漸消褪。紅、綠熒光在36h檢測后,信號(hào)完全消失。結(jié)論:1.我們成功構(gòu)建了小鼠活體內(nèi)細(xì)粒棘球蚴熒光模型,并可通過紅/綠熒光強(qiáng)度比來檢測原頭蚴活性。
[Abstract]:Objective: 1. to try to stain the Echinococcus granulosus in vitro, and to analyze the effect of different drugs on the activity of echinococcosis through the intensity of red green fluorescence signal..2. construction of the fluorescent luminescence model in the living body of Echinococcus granulosus mice, and the localization and quantitative analysis of the infection foci of Echinococcus granulosus in the liver of mice. Method: 1. aseptic separation and extraction of the original cercariae were divided into the control group, the metformin treatment group (10 m M metformin was added to the metarcariae medium) and the combined treatment group (10 m M metformin and 15 mu M albendazole (equivalent to 4.2 mu m L) were joined in the culture medium of the original cercariae). The original cercariae active.2. was stained with JC-1 fluorescent dyes for different treatment groups. The fluorescence intensity of the original cercariae was collected and recorded under confocal fluorescence microscope, and the fluorescence intensity of the primary cercariae in each group was recorded and recorded by.3.. The original cercariae of different treatment groups were placed in the 24 foramen plate, and the fluorescence intensity of the three groups of cercariae was continuously cultured and the fluorescent intensity of small animals was used for the fluorescence intensity of the cercariae. Dynamic monitoring was carried out until the fluorescent signal disappeared.4. and the original cercariae of different treatment groups were stained under the membrane of the CF-1 male mice liver. The mice in the different injection groups were placed in the small animal luminescence imaging instrument for dynamic imaging. The control group was selected to carry out the fluorescence ladder test, and the attenuation trend of the fluorescence signal was analyzed. The results were as follows: 1 Compared with the control group, the activity of the primary cercariae in the Met treatment group decreased to a certain extent. The activity of the ABZSO and Met drugs in the combined use group was the most significant decrease in the red fluorescence intensity of the original cercariae in the.2. control group, and the red fluorescence intensity of the original cercariae in the Met drug treatment group was not significantly different from the green fluorescence; ABZSO The green fluorescence intensity of the primary cercariae in the combination group with the Met drug was significantly stronger than that of the red fluorescent.3. three group. The decline trend of the combined drug group was the most obvious, followed by the metformin drug treatment group and the control group. In the metformin treatment group, the obvious enhancement trend was found, and the combination group was stronger than the metformin group, and then the decrease trend correspondingly appeared. The green fluorescence of the control group was at a lower level from the beginning of detection, and there was a certain decline trend. Compared with the control group, the experimental group had a significant statistical difference (P0.05).4. successfully planted. The red fluorescence intensity of the control group was significantly greater than that of green fluorescence in the control group, and the red fluorescence intensity was stronger than the green fluorescence. The red / green fluorescence intensity ratio of the 4.1. metformin group was compared with the green fluorescence. The red / green ratio was 1.2, and the red / green ratio was 1.2. The red fluorescence region of the combined drug group was less than the green fluorescence, and the red fluorescence intensity was also weaker than the green fluorescence. The red / green ratio was high in the early observation of the liver region of the 0.5. control group, and then gradually weakened. After 36h detection, the signal completely disappeared. Conclusion: 1. we successfully constructed the fluorescence model of Echinococcus granulosus in vivo, and can detect the activity of the echinococcosis by red / green fluorescence intensity ratio.

【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R532.32;R-332
，

本文編號(hào)：1780681