Metabolic Impact of 4-Hydroxynonenal on Macrophage-Like RAW 264.7 Function and Activation
Metabolic Impact of 4-Hydroxynonenal on Macrophage-Like RAW 264.7 Function and Activation
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DOI：
10.1021/tx3001048
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发表时间：
2012-08-01
影响因子：
4.1
通讯作者：
Wright, David W.
中科院分区：
文献类型：
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作者：
Harry, Reese S.;Hiatt, Leslie A.;Wright, David W.
暴露于4-羟基壬烯醛（HNE）后的巨噬细胞代谢反应的代谢谱表明，HNE不仅会抑制产生超氧化物的酶，而且还可能导致低场信号通路的损伤。采用多分析物微生理测量法（MAMP）同时测量细胞外酸化，乳酸产生和氧消耗的扰动，以检查有氧和厌氧途径。结合与佛波醇肉豆蔻酸酯乙酸酯（PMA）和HNE的免疫抑制的氧化爆发的激活，HNE毒性的复杂性质被确定为浓度和时间依赖性。进一步分析用于评估HNE对活性氧（ROS）产生和蛋白激酶C（PKC）的时间效应。随着PKC活性降低，HNE水平增加，表明PKC是HNE在氧化爆发前内收的靶点。此外，PKC定位到细胞膜被阻止与HNE的引入，证明了HNE加合对NADPH活化的后果。HNE对ROS的损伤表明HNE在泡沫细胞形成和组织损伤中的作用比已知的更大。虽然已经进行了工作，以了解特定的信号转导通路的HNE的调节的效果，关于其参与细胞代谢作为一个整体的细节通常是未知的。本研究探讨了HNE对巨噬细胞氧化爆发的影响，并确定PKC作为HNE抑制和最终代谢反应的关键蛋白。
Metabolic profiling of macrophage metabolic response upon exposure to 4-hydroxynonenal (HNE) demonstrates that HNE does not simply inactivate superoxide-generating enzymes but also could be responsible for the impairment of downfield signaling pathways. Multianalyte microphysiometry (MAMP) was employed to simultaneously measure perturbations in extracellular acidification, lactate production, and oxygen consumption for the examination of aerobic and anaerobic pathways. Combining the activation of oxidative burst with phorbol myristate acetate (PMA) and the immunosuppression with HNE, the complex nature of HNE toxicity was determined to be concentration- and time-dependent. Further analysis was utilized to assess the temporal effect of HNE on reactive oxygen species (ROS) production and on protein kinase C (PKC). Increased levels of HNE with decreasing PKC activity suggest that PKC is a target for HNE adductation prior to oxidative burst. Additionally, localization of PKC to the cell membrane was prevented with the introduction of HNE, demonstrating a consequence of HNE adductation on NADPH activation. The impairment of ROS by HNE suggests that HNE has a greater role in foam cell formation and tissue damage than is already known. Although work has been performed to understand the effect of HNE's regulation of specific signaling pathways, details regarding its involvement in cellular metabolism as a whole are generally unknown. This study examines the impact of HNE on macrophage oxidative burst and identifies PKC as a key protein for HNE suppression and eventual metabolic response.