Gupta Shakti, Kihara Yasuyuki, Maurya Mano R, Norris Paul C, Dennis Edward A, Subramaniam Shankar
Department of Bioengineering and San Diego Supercomputer Center, University of California at San Diego , 9500 Gilman Drive, La Jolla, California 92093-0412, United States.
Department of Chemistry and Biochemistry and Pharmacology, School of Medicine, University of California at San Diego , 9500 Gilman Drive, La Jolla, California 92093-0601, United States.
J Phys Chem B. 2016 Aug 25;120(33):8346-53. doi: 10.1021/acs.jpcb.6b02036. Epub 2016 Apr 26.
Arachidonic acid (AA), a representative ω6-polyunsaturated fatty acid (PUFA), is a precursor of 2-series prostaglandins (PGs) that play important roles in inflammation, pain, fever, and related disorders including cardiovascular diseases. Eating fish or supplementation with the ω3-PUFAs such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) is widely assumed to be beneficial in preventing cardiovascular diseases. A proposed mechanism for a cardio-protective role of ω3-PUFAs assumes competition between AA and ω3-PUFAs for cyclooxygenases (COX), leading to reduced production of 2-series PGs. In this study, we have used a systems biology approach to integrate existing knowledge and novel high-throughput data that facilitates a quantitative understanding of the molecular mechanism of ω3- and ω6-PUFA metabolism in mammalian cells. We have developed a quantitative computational model of the competitive metabolism of AA and EPA via the COX pathway through a two-step matrix-based approach to estimate the rate constants. This model was developed by using lipidomic data sets that were experimentally obtained from EPA-supplemented ATP-stimulated RAW264.7 macrophages. The resulting model fits the experimental data well for all metabolites and demonstrates that the integrated metabolic and signaling networks and the experimental data are consistent with one another. The robustness of the model was validated through parametric sensitivity and uncertainty analysis. We also validated the model by predicting the results from other independent experiments involving AA- and DHA-supplemented ATP-stimulated RAW264.7 cells using the parameters estimated with EPA. Furthermore, we showed that the higher affinity of EPA binding to COX compared with AA was able to inhibit AA metabolism effectively. Thus, our model captures the essential features of competitive metabolism of ω3- and ω6-PUFAs.
花生四烯酸(AA)是一种具有代表性的ω6 - 多不饱和脂肪酸(PUFA),是2 - 系列前列腺素(PGs)的前体,在炎症、疼痛、发热以及包括心血管疾病在内的相关病症中发挥着重要作用。人们普遍认为,食用鱼类或补充ω3 - 多不饱和脂肪酸,如二十碳五烯酸(EPA)和二十二碳六烯酸(DHA),对预防心血管疾病有益。ω3 - 多不饱和脂肪酸具有心脏保护作用的一种推测机制是,AA与ω3 - 多不饱和脂肪酸竞争环氧化酶(COX),导致2 - 系列PGs的生成减少。在本研究中,我们采用系统生物学方法整合现有知识和新的高通量数据,以促进对哺乳动物细胞中ω3 - 和ω6 - 多不饱和脂肪酸代谢分子机制的定量理解。我们通过基于矩阵的两步法估计速率常数,建立了AA和EPA通过COX途径竞争性代谢的定量计算模型。该模型是利用从补充了EPA的ATP刺激的RAW264.7巨噬细胞实验获得的脂质组学数据集开发的。所得模型对所有代谢物的实验数据拟合良好,表明整合的代谢和信号网络与实验数据相互一致。通过参数敏感性和不确定性分析验证了模型的稳健性。我们还通过使用用EPA估计的参数预测涉及补充了AA和DHA的ATP刺激的RAW264.7细胞的其他独立实验结果,对模型进行了验证。此外,我们表明,与AA相比,EPA与COX结合的亲和力更高,能够有效抑制AA代谢。因此,我们的模型捕捉到了ω3 - 和ω6 - 多不饱和脂肪酸竞争性代谢 的基本特征。