不同阶段ApoE-/-小鼠动脉粥样硬化炎症差异基因表达的比较研究  

Preliminary comparison of inflammatory differential gene expression during atherosclerosis in ApoE-/- mice

作　　者：于永慧 董瑞红[1,2] 刘剑刚 张大武[1] 王承龙[1] YU Yong-hui;DONG Rui-hong;LIU Jian-gang;ZHANG Da-wu;WANG Cheng-long(Cardiovascular Center, Xiyuan Hospital, China Academy of Chinese Medical Science, Beijing 100091, China;Beijing Mentougou Hospital of Traditional Chinese Medicine, Beijing 102300, China)

机构地区：[1]中国中医科学院西苑医院心血管病中心,北京100091 [2]北京市门头沟区中医医院,北京102300

出　　处：《中国病理生理杂志》2019年第9期1694-1699,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81273934)

摘　　要：目的:利用Agilent基因芯片筛查 ApoE -/-小鼠在动脉粥样硬化(AS)不同阶段(10周龄、15周龄和25周龄)炎症基因的表达谱变化,并利用RT-qPCR对筛选出的差异基因进行验证和演变分析。方法: 60只雄性 ApoE -/-小鼠随机分为3组,每组20只,设定10周龄为AS初期组,15周龄为AS早期组,25周龄为AS后期组,相同遗传背景的C57BL/6J野生型小鼠作为空白对照组。差异基因筛选采用Cy3标记的cRNA和基因芯片杂交,经洗脱、扫描图像、提取数据和标准化后,筛选出 P ≤0.05且上调或下调倍数≥2.0的差异基因。最后对筛选出的上述基因进行RT-qPCR定量分析。结果:(1)与对照组相比, ApoE -/-小鼠显著变化的差异基因为10周龄895个,15周龄540个,25周龄591个;(2)对差异基因功能进行GO条目分析,并去除与AS炎症反应无关的基因后选定白细胞介素12a(IL-12a)、基质金属肽酶12(MMP-12)、IL-1β、生长分化因子15(GDF-15)和干扰素γ(IFN-γ)为目标差异基因;(3)目标差异基因的RT-qPCR验证结果显示,与对照组比较, ApoE -/-小鼠10周龄IL-12a和MMP-12上调,IL-1β下调;15周龄GDF-15上调,IL-12a和IL-1β下调;25周龄IL-12a、MMP-12和GDF-15上调( P <0.05);并且10周龄IL-12a上调、15周龄IL-1β下调、25周龄MMP-12和GDF-15上调更为显著( P <0.01)。结论: AS炎症基因的动态变化过程为药物干预不同阶段AS提供了干预靶点。AIM: To screen the expression of inflammatory genes associated with atherosclerosis (AS) in different weeks of ApoE -/- mice using Agilent gene expression profile chip (AGEPC). METHODS: Male ApoE -/- mice ( n =60) were randomly divided into 3 groups: initial phase of AS (10 weeks old), early phase of AS (15 weeks old), and late phase of AS (25 weeks old). Homologous wild-type C57BL/6J mice were used for the control. The RNA samples of the arcus aortae from these mice were isolated. Total RNA from each sample was labeled with Cy3 and hybridized with AGEPC, and microarray detection was conducted. After washing, scaning, acquiring data, and standardized analysis, the expressed genes with default threshold of statistical significance of P ≤0.05 and fold change≥2.0 were selected. The expression of these genes were further verified by RT-qPCR. RESULTS: Compared with the control group, there were 895 differential genes in 10 weeks of ApoE -/- mice, while 540 genes in 15 weeks, and 591 genes in 25 weeks, respectively. KEGG pathway and gene ontology (GO) analyses revealed that those diversely expressed genes related to inflammation were particularly arresting. Several selected genes including interleukin-12a (IL-12a), matrix metallopeptidase-12 (MMP-12), IL-1β, growth differentiation factor-15 (GDF-15) and interferon-γ(IFN-γ) were validated by RT-qPCR. Compared with the control group, the expression levels of IL-12a and MMP-12 were up-regulated while IL-1β was down-regulated in 10 weeks, the expression level of GDF-15 was up-regulated while the IL-12a and IL-1β levels were down-regulated in 15 weeks, and the levels of IL-12a, MMP-12 and GDF-15 were up-regulated in 25 weeks ( P <0.05). Moreover, the increased level of IL-12a in 10 weeks, decreased level of IL-1β in 15 weeks, and increased levels of MMP-12 and GDF-15 in 25 weeks were even more statistically significant ( P <0.01). CONCLUSION: The changes of inflammatory gene expression in different phases of AS suggest an important direction for medical interventi

关 键 词：动脉粥样硬化 APOE -/-小鼠 基因芯片 炎症因子 

分 类 号：R543.12[医药卫生—心血管疾病] R363.2[医药卫生—内科学]