本文選題：泡球蚴　切入點(diǎn)：BALB/c小鼠　出處：《新疆醫(yī)科大學(xué)》2013年碩士論文

【摘要】：目的：初步篩選泡球蚴感染宿主肝臟中miRNAs的表達(dá)譜并探討miR-106b-25基因簇在肝泡型包蟲病中的表達(dá)及意義。方法：取雌性Balb/c小鼠隨機(jī)分為實(shí)驗(yàn)組、對(duì)照組和正常組，實(shí)驗(yàn)組小鼠腹腔注射泡球蚴，對(duì)照組腹腔注射等容量生理鹽水，正常組不作處理并在接種當(dāng)日采集肝臟標(biāo)本。接種后1月、3月和6月分別各取5只造模成功的實(shí)驗(yàn)組和對(duì)照組小鼠肝臟標(biāo)本，提取RNA；將感染3月小鼠肝臟的小分子RNA標(biāo)記后與miRNAs微陣列芯片雜交。掃描雜交后的芯片并進(jìn)行數(shù)據(jù)分析，得到肝泡型包蟲病的miRNAs表達(dá)譜。熒光定量RT-PCR法檢測(cè)在1月、3月、6月三個(gè)時(shí)間點(diǎn)泡球蚴病小鼠肝臟中miR-93、miR-106b、miR-494、miR-152、miR-191和miR-199a的相對(duì)表達(dá)，預(yù)測(cè)miR-93和miR-106b的靶基因并初步分析。結(jié)果：基因芯片結(jié)果顯示，相比對(duì)照組，，實(shí)驗(yàn)組小鼠肝臟組織中有46個(gè)差異表達(dá)的miRNAs，29個(gè)上調(diào)，17個(gè)下調(diào)；其中上調(diào)2倍以上的有19個(gè)，下調(diào)大于2倍的有2個(gè)。熒光定量RT-PCR驗(yàn)證結(jié)果：miR-93、miR-106b、miR-494、miR-152、miR-191和miR-199a在感染1月實(shí)驗(yàn)組小鼠肝內(nèi)的相對(duì)表達(dá)量與對(duì)照組和正常組比較差異無統(tǒng)計(jì)學(xué)意義（P0.05）。在感染3月實(shí)驗(yàn)組小鼠肝內(nèi)的相對(duì)表達(dá)量與對(duì)照組和正常組比較均顯著增高（P0.05）；而miR-494和miR-191在感染6月的相對(duì)表達(dá)量與對(duì)照組和正常組比較顯著增高（P0.05），miR-93、miR-106b、miR-152和miR-199a差異則無統(tǒng)計(jì)學(xué)意義（P0.05），與芯片結(jié)果一致。生物信息學(xué)分析發(fā)現(xiàn)，miR-93和miR-106b的共同靶基因p21可能與細(xì)胞凋亡及細(xì)胞周期調(diào)控等生物學(xué)功能相關(guān)。結(jié)論：泡球蚴感染后的宿主肝臟miRNAs表達(dá)譜發(fā)生明顯改變，提示miRNAs可能參與了泡球蚴肝損傷的過程，推測(cè)miR-106b-25基因簇可能通過下調(diào)靶基因p21參與其中。
[Abstract]:Objective: to screen the miRNAs expression profile of the host liver infected with hydatid alveolar echinococci and to explore the expression and significance of miR-106b-25 gene cluster in hepatic alveolar hydatid disease.Methods: female Balb/c mice were randomly divided into three groups: experimental group, control group and normal group. Mice in experimental group were injected intraperitoneally with alveolar hydatid and control group were injected with normal saline of equal volume. The normal group was not treated and the liver samples were collected on the day of inoculation.After inoculation for 1 month, 3 months and 6 months after inoculation, 5 mice liver samples of successful model making and control group were obtained, respectively, and miRNAs microarray microarrays were hybridized with small molecule RNA labeled with miRNAs.The miRNAs expression profile of hepatic alveolar hydatid disease was obtained by scanning the hybridization chip and analyzing the data.Fluorescence quantitative RT-PCR assay was used to detect the relative expression of miR-93r-106bmr-494miR-152 miR-191 and miR-199a in the liver of mice with alveolar echinococcosis at three time points in January, March and June. The target genes of miR-93 and miR-106b were predicted and preliminarily analyzed.Results: compared with the control group, there were 46 differentially expressed miRNAss in the liver of the experimental group, 29 upregulated and 17 down-regulated, of which 19 were more than 2 times up-regulated and 2 more down-regulated.The results of fluorescence quantitative RT-PCR test showed that there was no significant difference in the relative expression of miR-494 miR-152 miR-191 and miR-199a between the control group and the control group.The relative expression of miR-494 and miR-191 in the liver of the experimental group was significantly higher than that of the control group and the normal group, while the relative expression of miR-494 and miR-191 in the 6th month of infection was significantly higher than that in the control group and the normal group. However, there was no difference in the expression of miR-494 and miR-191 between the control group and the normal group.The statistical significance was P0.05, which was consistent with the result of the chip.Bioinformatics analysis revealed that the common target gene p21 of miR-93 and miR-106b may be related to cell apoptosis and cell cycle regulation and other biological functions.Conclusion: the expression profile of miRNAs in the host liver after infection of alveolar hydatid is obviously changed, suggesting that miRNAs may be involved in the process of hepatic injury of alveolar hydatid, and it is speculated that miR-106b-25 gene cluster may participate in it by down-regulation of target gene p21.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R532.32

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 劉真;賈赤宇;胡永亮;毛旭虎;鄒全明;;幽門螺桿菌感染誘導(dǎo)microRNA-146a表達(dá)的機(jī)制研究[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2011年24期

2 馬兆龍;楊煉;陳立波;黃金明;王冬冬;王國(guó)斌;;miRNA在HBV從感染經(jīng)由肝硬化到肝癌進(jìn)程中表達(dá)譜的變化[J];世界華人消化雜志;2009年20期

3 魏緒法;邵英梅;王俊華;李瑤;張傳山;劉輝;姜濤;盧曉梅;溫浩;林仁勇;;泡球蚴感染中期小鼠肝臟病灶周圍肉芽腫TGF-β1及其受體TβRⅠ和p-Smad2/3的表達(dá)及意義[J];中國(guó)病原生物學(xué)雜志;2010年12期

4 王俊華;魏緒法;張傳山;呂國(guó)棟;姜濤;盧曉梅;溫浩;林仁勇;;Ⅰ、Ⅲ、Ⅳ膠原基因在泡球蚴感染中期小鼠肝臟中的表達(dá)及意義[J];中國(guó)病原生物學(xué)雜志;2012年09期

本文編號(hào)：1708717