小鼠肝泡型包虫病中miRNAs表达谱的初步筛选及miR-106b-25基因簇的表达和意义

发布时间：2018-04-04 06:14

本文选题：泡球蚴　切入点：BALB/c小鼠　出处：《新疆医科大学》2013年硕士论文

【摘要】：目的：初步筛选泡球蚴感染宿主肝脏中miRNAs的表达谱并探讨miR-106b-25基因簇在肝泡型包虫病中的表达及意义。方法：取雌性Balb/c小鼠随机分为实验组、对照组和正常组，实验组小鼠腹腔注射泡球蚴，对照组腹腔注射等容量生理盐水，正常组不作处理并在接种当日采集肝脏标本。接种后1月、3月和6月分别各取5只造模成功的实验组和对照组小鼠肝脏标本，提取RNA；将感染3月小鼠肝脏的小分子RNA标记后与miRNAs微阵列芯片杂交。扫描杂交后的芯片并进行数据分析，得到肝泡型包虫病的miRNAs表达谱。荧光定量RT-PCR法检测在1月、3月、6月三个时间点泡球蚴病小鼠肝脏中miR-93、miR-106b、miR-494、miR-152、miR-191和miR-199a的相对表达，预测miR-93和miR-106b的靶基因并初步分析。结果：基因芯片结果显示，相比对照组，，实验组小鼠肝脏组织中有46个差异表达的miRNAs，29个上调，17个下调；其中上调2倍以上的有19个，下调大于2倍的有2个。荧光定量RT-PCR验证结果：miR-93、miR-106b、miR-494、miR-152、miR-191和miR-199a在感染1月实验组小鼠肝内的相对表达量与对照组和正常组比较差异无统计学意义（P0.05）。在感染3月实验组小鼠肝内的相对表达量与对照组和正常组比较均显著增高（P0.05）；而miR-494和miR-191在感染6月的相对表达量与对照组和正常组比较显著增高（P0.05），miR-93、miR-106b、miR-152和miR-199a差异则无统计学意义（P0.05），与芯片结果一致。生物信息学分析发现，miR-93和miR-106b的共同靶基因p21可能与细胞凋亡及细胞周期调控等生物学功能相关。结论：泡球蚴感染后的宿主肝脏miRNAs表达谱发生明显改变，提示miRNAs可能参与了泡球蚴肝损伤的过程，推测miR-106b-25基因簇可能通过下调靶基因p21参与其中。
[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.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R532.32

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