Cholesterol loading augments oxidative stress in macrophages
Cholesterol loading augments oxidative stress in macrophages
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DOI：
10.1016/j.febslet.2005.12.102
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发表时间：
2006-02-06
期刊：
影响因子：
3.5
通讯作者：
Huang, GS
中科院分区：
文献类型：
--
作者：
Hung, YC;Hong, MY;Huang, GS
为了研究将游离胆固醇加载到巨噬细胞中的分子后果，我们进行了大规模的基因表达研究，以分析由人THP-1细胞系诱导的乙酰化LDL加载的泡沫细胞（AFC）和氧化LDL加载的泡沫细胞（OFC）。利用9600个基因芯片数据集进行聚类分析。AFC和OFC共享共同的表达谱，然而，这两种治疗之间存在足够的差异，AFC和OFC呼吁作为两个独立的实体。我们鉴定了AFC和OFC中80个上调基因和48个下调基因。差异表达基因的功能注释表明，细胞凋亡，细胞外基质，氧化应激和细胞增殖失调。我们还确定了87个差异表达的基因独特的AFC和31个基因的OFC。AFC的独特表达基因与激酶活性、ATP结合活性和转运蛋白活性相关，而OFC的独特表达基因与细胞信号传导和粘附相关。为了验证氧化应激是AFC和OFC的共同特征的假设，我们使用与氧化应激相关的基因进行了聚类分析，但我们无法以这种方式区分AFC和OFC。采用实时荧光定量PCR和ELISA方法检测泡沫细胞中白细胞介素1 β（IL 1 β）的转录和分泌蛋白。在泡沫细胞中迅速诱导IL 1 β，但对于AFC，RNA水平和蛋白水平立即下降并减弱。为了检测泡沫细胞中活性氧的水平，我们进行了氢乙啶染色，并观察到高水平的超氧阴离子。我们的结论是，加载游离胆固醇会诱导高水平的超氧阴离子，增加氧化应激，并引发巨噬细胞中的短暂炎症反应。(c)2006年由Elsevier B. V.代表欧洲生物化学学会联合会出版。
To investigate the molecular consequence of loading free cholesterol into macrophages, we conducted a large-scale gene expression study to analyze acetylated-LDL-laden foam cells (AFC) and oxidized-LDL-laden foam cells (OFC) induced from human THP-1 cell lines. Cluster analysis was performed using 9600-gene microarray datasets from time course experiment. AFC and OFC shared common expression profiles; however, there were sufficient differences between these two treatments that AFC and OFC appealed as two separate entities. We identified 80 commonly upregulated genes and 48 commonly downregulated genes in AFC and OFC. Functional annotation of the differentially expressed genes indicated that apoptosis, extracellular matrix, oxidative stress, and cell proliferation was deregulated. We also identified 87 differentially expressed genes unique for AFC and 31 genes for OFC. The uniquely expressed genes of AFC are associated with kinase activity, ATP binding activity, and transporter activity, while unique genes for OFC are associated with cell signaling and adhesion. To validate the hypothesis that oxidative stress is a common feature for AFC and OFC, we performed a cluster analysis employing the genes related to oxidative stress, but we were unable to distinguish AFC from OFC in this manner. We performed real-time RTPCR and ELISA on foam cells to examine the transcripts and secreted protein of interleukin 1 beta (IL1 beta). IL1 beta was rapidly induced in foam cells, but for AFC both RNA level and protein level dropped immediately and was attenuated. To detect levels of reactive oxygen species in foam cells we conducted hydroethidine staining and observed high levels of superoxide anion. We conclude that loading free cholesterol induces high levels of superoxide anion, increases oxidative stress, and triggers a transient inflammatory response in macrophages. (c) 2006 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.