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RGD‐peptide lunasin inhibits Akt‐mediated NF‐κB activation in human macrophages through interaction with the αVβ3 integrin
Author(s) -
Cam Anthony,
de Mejia Elvira Gonzalez
Publication year - 2012
Publication title -
molecular nutrition and food research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.495
H-Index - 131
eISSN - 1613-4133
pISSN - 1613-4125
DOI - 10.1002/mnfr.201200301
Subject(s) - protein kinase b , proinflammatory cytokine , tumor necrosis factor alpha , chemistry , inflammation , nitric oxide synthase , biology , microbiology and biotechnology , nitric oxide , signal transduction , biochemistry , immunology , endocrinology
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‐κB)‐dependent markers of inflammation and to characterize the physical interaction of lunasin with the αVβ3 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 μM) 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‐κB p65 by 59.5 and 74.5%, respectively. Lunasin (50 μM) reduced exogenous release of prostaglandin E 2 and tumor necrosis factor‐α by 92.5 and 94.9%, respectively. Vitronectin (10 μg/mL), an integrin ligand, increased expression of proinflammatory markers, whereas lunasin (50 μM) attenuated them. Co‐IP of lunasin‐treated cells confirmed direct interaction with αVβ3 integrin and LC/MS/MS verified its identity. Lunasin was detected within intracellular vesicles and reduced total αVβ3 intensity as observed by fluorescence microscopy. Conclusion Lunasin inhibited αVβ3 integrin‐mediated proinflammatory markers and downregulated Akt‐mediated NF‐κB pathways through interaction with αVβ3 integrin.