RGD-peptide lunasin inhibits Akt-mediated NF-?B activation in human macrophages through interaction with the αVβ3 integrin
RGD-peptide lunasin inhibits Akt-mediated NF-?B activation in human macrophages through interaction with the αVβ3 integrin
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DOI：
10.1002/mnfr.201200301
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发表时间：
2012-10-01
影响因子：
5.2
通讯作者：
de Mejia, Elvira Gonzalez
中科院分区：
文献类型：
--
作者：
Cam, Anthony;de Mejia, Elvira Gonzalez
在美国，心血管疾病是导致死亡的主要原因，在炎症条件下调节异常的巨噬细胞活性对其预防至关重要。目的是确定lunasin对Akt介导的核因子-κ B（NF-？B）-依赖性炎症标志物，并表征lunasin与脂多糖（LPS）-诱导的人THP-1巨噬细胞中α V β 3整联蛋白受体的物理相互作用。方法和结果使用免疫测定、免疫共沉淀（Co-IP）和荧光共聚焦显微镜在LPS诱导的THP-1人巨噬细胞中体外评估lunasin的作用。Lunasin（50 μ M）减少环氧合酶-2，诱导型一氧化氮合酶和NO水平分别为57.9%，64.5%和76.2%，并抑制磷酸化Akt和NF-？B p65分别降低59.5%和74.5%。Lunasin（50 μ M）减少外源性释放的前列腺素E2和肿瘤坏死因子-α分别为92.5%和94.9%。玻连蛋白（10 μ g/mL），一种整合素配体，增加了促炎标志物的表达，而lunasin（50 μ M）减弱了它们。Iunasin处理的细胞的Co-IP证实了与aV β 3整联蛋白的直接相互作用，LC/MS/MS验证了其身份。在胞内囊泡内检测到Lunasin，并且如通过荧光显微镜观察到的，降低了总aV β 3强度。结论Lunasin可抑制aV β 3整合素介导的促炎性标志物，下调Akt介导的NF-？B途径通过与α V β 3整联蛋白相互作用。
Scope Cardiovascular disease is the leading cause of mortality in the United States and regulation of aberrant macrophage activity under inflammatory conditions is critical for its prevention. The objective was to determine the effect of lunasin on the inhibition of Akt-mediated activation of nuclear factor-kappa B (NF-?B)-dependent markers of inflammation and to characterize the physical interaction of lunasin with the aV beta 3 integrin receptor in lipopolysaccharide (LPS)-induced human THP-1 macrophages. Methods and results The effect of lunasin was evaluated in vitro in LPS-induced THP-1 human macrophages using immunoassays, co-immunoprecipitation (Co-IP), and fluorescence confocal microscopy. Lunasin (50 mu M) reduced cyclooxygenase-2, inducible nitric oxide synthase, and NO levels by 57.9, 64.5, and 76.2%, respectively, and inhibited the activation of phosphorylated Akt and NF-?B p65 by 59.5 and 74.5%, respectively. Lunasin (50 mu M) reduced exogenous release of prostaglandin E2 and tumor necrosis factor-a by 92.5 and 94.9%, respectively. Vitronectin (10 mu g/mL), an integrin ligand, increased expression of proinflammatory markers, whereas lunasin (50 mu M) attenuated them. Co-IP of lunasin-treated cells confirmed direct interaction with aV beta 3 integrin and LC/MS/MS verified its identity. Lunasin was detected within intracellular vesicles and reduced total aV beta 3 intensity as observed by fluorescence microscopy. Conclusion Lunasin inhibited aV beta 3 integrin-mediated proinflammatory markers and downregulated Akt-mediated NF-?B pathways through interaction with aV beta 3 integrin.