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Preliminary comparison of inflammatory differential gene expression during atherosclerosis in ApoE-/- mice
Authors:YU Yong-hui  DONG Rui-hong  LIU Jian-gang  ZHANG Da-wu  WANG Cheng-long
Institution:1. Cardiovascular Center, Xiyuan Hospital, China Academy of Chinese Medical Science, Beijing 100091, China; 2. Beijing Mentougou Hospital of Traditional Chinese Medicine, Beijing 102300, China
Abstract: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 intervention of AS.
Keywords:Atherosclerosis  ApoE-/- mice  Gene chip  Inflammatory factors  
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