A novel role of sesamol in inhibiting NF-κB-mediated signaling in platelet activation

Journal of Biomedical Science

Joen-Rong Sheu²  Shih-Yi Huang¹  Song-Chow Lin²  Cheng-Wen Chiang³  Eng-Thiam Ong³  Wan-Jung Lu²  Chao-Chien Chang³ 

[1] School of Nutrition and Health Sciences, Taipei Medical University, 250 Wu-Hsing St., Taipei 11031, Taiwan;Department of Pharmacology, School of Medicine, Taipei Medical University, 250 Wu-Hsing St., Taipei 11031, Taiwan;Department of Cardiology, Cathay General Hospital, 280 Renai Rd. Sec.4, Taipei 10630, Taiwan

关键词: sesamol;    platelet activation;    protein kinase A;    intracellular Ca²⁺;    IKK;    IκBα;   

Others  :  1146728
DOI  :  10.1186/1423-0127-18-93

 received in 2011-08-12, accepted in 2011-12-14,  发布年份 2011

【 摘 要 】

Background

Platelet activation is relevant to a variety of coronary heart diseases. Our previous studies revealed that sesamol possesses potent antiplatelet activity through increasing cyclic AMP formation. Although platelets are anucleated cells, they also express the transcription factor, NF-κB, that may exert non-genomic functions in platelet activation. Therefore, we further investigated the inhibitory roles of sesamol in NF-κB-mediated platelet function.

Methods

Platelet aggregation, Fura 2-AM fluorescence, and immunoblotting analysis were used in this study.

Results

NF-κB signaling events, including IKKβ phosphorylation, IκBα degradation, and p65 phosphorylation, were markedly activated by collagen (1 μg/ml) in washed human platelets, and these signaling events were attenuated by sesamol (2.5~25 μM). Furthermore, SQ22536 and ODQ, inhibitors of adenylate cyclase and guanylate cyclase, respectively, strongly reversed the sesamol (25 μM)-mediated inhibitory effects of IKKβ phosphorylation, IκBα degradation, and p65 phosphorylation stimulated by collagen. The protein kinase A (PKA) inhibitor, H89, also reversed sesamol-mediated inhibition of IκBα degradation. Moreover, BAY11-7082, an NF-κB inhibitor, abolished IκBα degradation, phospholipase C (PLC)γ2 phosphorylation, protein kinase C (PKC) activation, [Ca²⁺]i mobilization, and platelet aggregation stimulated by collagen. Preincubation of platelets with the inhibitors, SQ22536 and H89, both strongly reversed sesamol-mediated inhibition of platelet aggregation and [Ca²⁺]i mobilization.

Conclusions

Sesamol activates cAMP-PKA signaling, followed by inhibition of the NF-κB-PLC-PKC cascade, thereby leading to inhibition of [Ca²⁺]i mobilization and platelet aggregation. Because platelet activation is not only linked to hemostasis, but also has a relevant role in inflammation and metastasis, our data demonstrating that inhibition of NF-κB interferes with platelet function may have a great impact when these types of drugs are considered for the treatment of cancer and various inflammatory diseases.

【 授权许可】

   
2011 Chang et al; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Hou YC, Tsai SY, Liu IL, Yu CP, Chao PD: Metabolic transformation of sesamol and ex vivo effect on 2,2'-azo-bis(2-amidinopropane)dihydrochloride-induced hemolysis. J Agric Food Chem 2008, 56:9636-9640.
• [2]Kaur IP, Saini A: Sesamol exhibits antimutagenic activity against oxygen species mediated mutagenicity. Mutat Res 2000, 470:71-76.
• [3]Zimmet P, Alberti KG, Shaw J: Global and societal implications of the diabetes epidemic. Nature 2001, 414:782-787.
• [4]Kapadia GJ, Azuine MA, Tokuda H, Takasaki M, Mukainaka T, Konoshima T, Nishino H: Chemopreventive effect of resveratrol, sesamol, sesame oil and sunflower oil in the Epstein-Barr virus early antigen activation assay and the mouse skin two-stage carcinogenesis. Pharmacol Res 2002, 45:499-505.
• [5]Sharma S, Kaur IP: Development and evaluation of sesamol as an antiaging agent. Int J Dermatol 2006, 45:200-208.
• [6]Jacklin A, Ratledge C, Welham K, Bilko D, Newton CJ: The sesame seed oil constituent, sesamol, induces growth arrest and apoptosis of cancer and cardiovascular cells. Ann N Y Acad Sci 2003, 1010:374-380.
• [7]Chen PR, Lee CC, Chang H, Tsai CE: Sesamol regulates plasminogen activator gene expression in cultured endothelial cells: a potential effect on the fibrinolytic system. J Nutr Biochem 2005, 16:59-64.
• [8]Chen PR, Tsai CE, Chang H, Liu TL, Lee CC: Sesamol induces nitric oxide release from human umbilical vein endothelial cells. Lipids 2005, 40:955-961.
• [9]Sheu JR, Hung WC, Wu CH, Ma MC, Kan YC, Lin CH, Lin MS, Luk HN, Yen MH: Reduction in lipopolysaccharide-induced thrombocytopenia by triflavin in a rat model of septicemia. Circulation 1999, 99:3056-3062.
• [10]Sheu JR, Lee CR, Lin CH, Hsiao G, Ko WC, Chen YC, Yen MH: Mechanisms involved in the antiplatelet activity of Staphylococcus aureus lipoteichoic acid in human platelets. Thromb Haemost 2000, 83:777-784.
• [11]Chang CC, Lu WJ, Chiang CW, Jayakumar T, Ong ET, Hsiao G, Fong TH, Chou DS, Sheu JR: Potent antiplatelet activity of sesamol in an in vitro and in vivo model: pivotal roles of cyclic AMP and p38 mitogen-activated protein kinase. J Nutr Biochem 2010, 21:1214-1221.
• [12]Ghosh S, Hayden MS: New regulators of NF-kappaB in inflammation. Nat Rev Immunol 2008, 8:837-848.
• [13]Liu F, Morris S, Epps J, Carroll R: Demonstration of an activation regulated NF-kappaB/I-kappaBalpha complex in human platelets. Thromb Res 2002, 106:199-203.
• [14]Schwarz UR, Walter U, Eigenthaler M: Taming platelets with cyclic nucleotides. Biochem Pharmacol 2001, 62:1153-1161.
• [15]Singer WD, Brown HA, Sternweis PC: Regulation of eukaryotic phosphatidylinositol-specific phospholipase C and phospholipase D. Annu Rev Biochem 1997, 66:475-509.
• [16]Pierce JW, Schoenleber R, Jesmok G, Best J, Moore SA, Collins T, Gerritsen ME: Novel inhibitors of cytokine-induced IkappaBalpha phosphorylation and endothelial cell adhesion molecule expression show anti-inflammatory effects in vivo. J Biol Chem 1997, 272:21096-21103.
• [17]Khetawat G, Faraday N, Nealen ML, Vijayan KV, Bolton E, Noga SJ, Bray PF: Human megakaryocytes and platelets contain the estrogen receptor beta and androgen receptor (AR): testosterone regulates AR expression. Blood 2000, 95:2289-2296.
• [18]Moraes LA, Paul-Clark MJ, Rickman A, Flower RJ, Goulding NJ, Perretti M: Ligand-specific glucocorticoid receptor activation in human platelets. Blood 2005, 106:4167-4175.
• [19]Ali FY, Davidson SJ, Moraes LA, Traves SL, Paul-Clark M, Bishop-Bailey D, Warner TD, Mitchell JA: Role of nuclear receptor signaling in platelets: antithrombotic effects of PPARbeta. FASEB J 2006, 20:326-328.
• [20]Malaver E, Romaniuk MA, D'Atri LP, Pozner RG, Negrotto S, Benzadon R, Schattner M: NF-kappaB inhibitors impair platelet activation responses. J Thromb Haemost 2009, 7:1333-1343.
• [21]Bunney TD, Katan M: Phospholipase C epsilon: linking second messengers and small GTPases. Trends Cell Biol 2006, 16:640-648.
• [22]Ragab A, Severin S, Gratacap MP, Aguado E, Malissen M, Jandrot-Perrus M, Malissen B, Ragab-Thomas J, Payrastre B: Roles of the C-terminal tyrosine residues of LAT in GPVI-induced platelet activation: insights into the mechanism of PLC gamma 2 activation. Blood 2007, 110:2466-2474.
• [23]Mangin P, Yuan Y, Goncalves I, Eckly A, Freund M, Cazenave JP, Gachet C, Jackson SP, Lanza F: Signaling role for phospholipase C gamma 2 in platelet glycoprotein Ib alpha calcium flux and cytoskeletal reorganization. Involvement of a pathway distinct from FcR gamma chain and Fc gamma RIIA. J Biol Chem 2003, 278:32880-32891.
• [24]Pascale A, Amadio M, Govoni S, Battaini F: The aging brain, a key target for the future: the protein kinase C involvement. Pharmacol Res 2007, 55:560-569.
• [25]Francis SH, Corbin JD: Structure and function of cyclic nucleotide-dependent protein kinases. Annu Rev Physiol 1994, 56:237-272.
• [26]Profirovic J, Gorovoy M, Niu J, Pavlovic S, Voyno-Yasenetskaya T: A novel mechanism of G protein-dependent phosphorylation of vasodilator-stimulated phosphoprotein. J Biol Chem 2005, 280:32866-32876.
• [27]Gambaryan S, Kobsar A, Rukoyatkina N, Herterich S, Geiger J, Smolenski A, Lohmann SM, Walter U: Thrombin and collagen induce a feedback inhibitory signaling pathway in platelets involving dissociation of the catalytic subunit of protein kinase A from an NFkappaB-IkappaB complex. J Biol Chem 2010, 285:18352-18363.