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Endocrine, Metabolic & Immune Disorders - Drug Targets

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ISSN (Print): 1871-5303
ISSN (Online): 2212-3873

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General Research Article

mRNA Expression Profile of SFKs and Involvement of SFKs in the Regulation of LPS-Induced Erk1/2 Signaling in PBMCs of Active BD Patients

Author(s): Sevgi Irtegun-Kandemir*, Irmak Icen-Taskin, Mehtap Bozkurt and Sevgi Kalkanli-Tas

Volume 19, Issue 6, 2019

Page: [809 - 817] Pages: 9

DOI: 10.2174/1871530319666190119101756

Price: $65

Abstract

Background: Behcet’s Disease (BD) is a multisystemic inflammatory disorder affecting large vessels, lungs joints, gastrointestinal and neurological systems. The pathogenesis of BD remains poorly understood. Identifying the key signaling pathway is crucial for a complete understanding of the pathogenesis of BD.

Objective: The aim of this study was to determine mRNA expression level of Src family kinases (SFKs) members and their involvement in lipopolysaccharide (LPS)-induced mitogen-activated protein kinases (MAPKs) regulation in peripheral blood mononuclear cells (PBMCs) of active BD patients.

Methods: Twenty- five active BD patients and twenty-five healthy controls were included in the study. PBMCs were isolated from total blood by density gradient centrifugation. The mRNA expression levels of SFKs members were measured by real-time quantitative PCR (RT-qPCR). The effect of SFKs activity on LPS-induced activation MAPKs (Erk1/2, p38 and JNK) was examined by Western blot.

Results: The mRNA expression levels of Hck, Src, Lyn, Yes and Fyn were found to be slightly decreased in active BD patients compared to the control subjects, but a slight change in mRNA level of SFKs members did not impact on protein levels and protein activity. LPS-induced Erk1/2 phosphorylation was significantly increased in the absence of SFKs activity in active BD patients. However, inhibition of SFKs activity had no effect on LPS-induced phosphorylation of p38 and JNK in both controls and active BD patients.

Conclusion: SFKs downregulate LPS-induced Erk1/2 phosphorylation in PBMCs of active BD patients.

Keywords: Behçet's disease, LPS, Mitogen-activated protein kinase, Src family kinases, PBMCs, TLR4.

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[1]
Yazici, H. The lumps and bumps of Behçet’s syndrome. Autoimmun. Rev., 2004, 3(Suppl. 1), S53-S54.
[PMID: 15309796]
[2]
Alpsoy, E. Behçet’s disease: A comprehensive review with a focus on epidemiology, etiology and clinical features, and management of mucocutaneous lesions. J. Dermatol., 2016, 43(6), 620-632.
[http://dx.doi.org/10.1111/1346-8138.13381] [PMID: 27075942]
[3]
Mat, M.C.; Sevim, A.; Fresko, I.; Tüzün, Y. Behçet’s disease as a systemic disease. Clin. Dermatol., 2014, 32(3), 435-442.
[http://dx.doi.org/10.1016/j.clindermatol.2013.11.012] [PMID: 24767193]
[4]
Verity, D.H.; Marr, J.E.; Ohno, S.; Wallace, G.R.; Stanford, M.R. Behçet’s disease, the Silk Road and HLA-B51: historical and geographical perspectives. Tissue Antigens, 1999, 54(3), 213-220.
[http://dx.doi.org/10.1034/j.1399-0039.1999.540301.x] [PMID: 10519357]
[5]
Mazzoccoli, G.; Matarangolo, A.; Rubino, R.; Inglese, M.; De Cata, A. Behçet syndrome: from pathogenesis to novel therapies. Clin. Exp. Med., 2016, 16(1), 1-12.
[http://dx.doi.org/10.1007/s10238-014-0328-z] [PMID: 25447032]
[6]
Houman, H.; Hamzaoui, A.; Ben Ghorbal, I.; Khanfir, M.; Feki, M.; Hamzaoui, K. Abnormal expression of chemokine receptors in Behçet’s disease: relationship to intracellular Th1/Th2 cytokines and to clinical manifestations. J. Autoimmun., 2004, 23(3), 267-273.
[http://dx.doi.org/10.1016/j.jaut.2004.07.005] [PMID: 15501397]
[7]
Mesquida, M.; Molins, B.; Llorenç, V.; Sainz de la Maza, M.; Hernandez, M.V.; Espinosa, G.; Adán, A. Proinflammatory cytokines and C-reactive protein in uveitis associated with Behçet’s disease. Mediators Inflamm., 2014.2014396204
[http://dx.doi.org/10.1155/2014/396204] [PMID: 24994946]
[8]
Faure, E.; Equils, O.; Sieling, P.A.; Thomas, L.; Zhang, F.X.; Kirschning, C.J.; Polentarutti, N.; Muzio, M.; Arditi, M. Bacterial lipopolysaccharide activates NF-kappaB through toll-like receptor 4 (TLR-4) in cultured human dermal endothelial cells. Differential expression of TLR-4 and TLR-2 in endothelial cells. J. Biol. Chem., 2000, 275(15), 11058-11063.
[http://dx.doi.org/10.1074/jbc.275.15.11058] [PMID: 10753909]
[9]
Lee, M.S.; Kim, Y.J. Signaling pathways downstream of pattern-recognition receptors and their cross talk. Annu. Rev. Biochem., 2007, 76, 447-480.
[http://dx.doi.org/10.1146/annurev.biochem.76.060605.122847] [PMID: 17328678]
[10]
Jury, E.C.; Kabouridis, P.S.; Abba, A.; Mageed, R.A.; Isenberg, D.A. Increased ubiquitination and reduced expression of LCK in T lymphocytes from patients with systemic lupus erythematosus. Arthritis Rheum., 2003, 48(5), 1343-1354.
[http://dx.doi.org/10.1002/art.10978] [PMID: 12746907]
[11]
Matache, C.; Onu, A.; Stefanescu, M.; Tanaseanu, S.; Dragomir, C.; Dolganiuc, A.; Szegli, G. Dysregulation of p56lck kinase in patients with systemic lupus erythematosus. Autoimmunity, 2001, 34(1), 27-38.
[http://dx.doi.org/10.3109/08916930108994123] [PMID: 11681490]
[12]
Clements, J.L.; Wolfe, J.; Cooper, S.M.; Budd, R.C. Reversal of hyporesponsiveness in lpr CD4-CD8- T cells is achieved by induction of cell cycling and normalization of CD2 and p59fyn expression. Eur. J. Immunol., 1994, 24(3), 558-565.
[http://dx.doi.org/10.1002/eji.1830240310] [PMID: 7510235]
[13]
Gong, P.; Angelini, D.J.; Yang, S.; Xia, G.; Cross, A.S.; Mann, D.; Bannerman, D.D.; Vogel, S.N.; Goldblum, S.E. TLR4 signaling is coupled to SRC family kinase activation, tyrosine phosphorylation of zonula adherens proteins, and opening of the paracellular pathway in human lung microvascular endothelia. J. Biol. Chem., 2008, 283(19), 13437-13449.
[http://dx.doi.org/10.1074/jbc.M707986200] [PMID: 18326860]
[14]
Irtegun, S.; Pektanc, G.; Akkurt, Z.M.; Bozkurt, M.; Turkcu, F.M.; Kalkanli-Tas, S. Pharmacological Inactivation of Src Family Kinases Inhibits LPS-Induced TNF-α Production in PBMC of Patients with Behçet’s Disease. Mediators Inflamm., 2016.20165414369
[http://dx.doi.org/10.1155/2016/5414369] [PMID: 27445436]
[15]
Smolinska, M.J.; Horwood, N.J.; Page, T.H.; Smallie, T.; Foxwell, B.M.J. Chemical inhibition of Src family kinases affects major LPS-activated pathways in primary human macrophages. Mol. Immunol., 2008, 45(4), 990-1000.
[http://dx.doi.org/10.1016/j.molimm.2007.07.026] [PMID: 17875324]
[16]
Thobe, B.M.; Frink, M.; Choudhry, M.A.; Schwacha, M.G.; Bland, K.I.; Chaudry, I.H. Src family kinases regulate p38 MAPK-mediated IL-6 production in Kupffer cells following hypoxia. Am. J. Physiol. Cell Physiol., 2006, 291(3), C476-C482.
[http://dx.doi.org/10.1152/ajpcell.00076.2006] [PMID: 16571868]
[17]
Hu, X.; Wu, X.; Xu, J.; Zhou, J.; Han, X.; Guo, J. Src kinase up-regulates the ERK cascade through inactivation of protein phosphatase 2A following cerebral ischemia. BMC Neurosci., 2009, 10(74), 74.
[http://dx.doi.org/10.1186/1471-2202-10-74] [PMID: 19602257]
[18]
Yoshizumi, M.; Abe, J.; Haendeler, J.; Huang, Q.; Berk, B.C. Src and Cas mediate JNK activation but not ERK1/2 and p38 kinases by reactive oxygen species. J. Biol. Chem., 2000, 275(16), 11706-11712.
[http://dx.doi.org/10.1074/jbc.275.16.11706] [PMID: 10766791]
[19]
Kim, S.R.; Jung, Y.R.; Kim, D.H.; An, H.J.; Kim, M.K.; Kim, N.D.; Chung, H.Y. Caffeic acid regulates LPS-induced NF-κB activation through NIK/IKK and c-Src/ERK signaling pathways in endothelial cells. Arch. Pharm. Res., 2014, 37(4), 539-547.
[http://dx.doi.org/10.1007/s12272-013-0211-6] [PMID: 23888332]
[20]
Napolitani, G.; Bortoletto, N.; Racioppi, L.; Lanzavecchia, A.; D’Oro, U. Activation of src-family tyrosine kinases by LPS regulates cytokine production in dendritic cells by controlling AP-1 formation. Eur. J. Immunol., 2003, 33(10), 2832-2841.
[http://dx.doi.org/10.1002/eji.200324073] [PMID: 14515267]
[21]
Criteria for diagnosis of Behçet’s disease. Lancet, 1990, 335(8697), 1078-1080.
[PMID: 1970380]
[22]
Livak, K.J.; Schmittgen, T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 2001, 25(4), 402-408.
[http://dx.doi.org/10.1006/meth.2001.1262] [PMID: 11846609]
[23]
Marotte, H.; Ahmed, S.; Ruth, J.H.; Koch, A.E. Blocking ERK-1/2 reduces tumor necrosis factor alpha-induced interleukin-18 bioactivity in rheumatoid arthritis synovial fibroblasts by induction of interleukin-18 binding protein A. Arthritis Rheum., 2010, 62(3), 722-731.
[http://dx.doi.org/10.1002/art.27269] [PMID: 20131228]
[24]
Amin, M.A.; Rabquer, B.J.; Mansfield, P.J.; Ruth, J.H.; Marotte, H.; Haas, C.S.; Reamer, E.N.; Koch, A.E. Interleukin 18 induces angiogenesis in vitro and in vivo via Src and Jnk kinases. Ann. Rheum. Dis., 2010, 69(12), 2204-2212.
[http://dx.doi.org/10.1136/ard.2009.127241] [PMID: 20679476]
[25]
Taheri, S.; Borlu, M.; Evereklioglu, C.; Ozdemir, S.Y.; Ozkul, Y. mRNA Expression Level of Interleukin Genes in the Determining Phases of Behçet’s Disease. Ann. Dermatol., 2015, 27(3), 291-297.
[http://dx.doi.org/10.5021/ad.2015.27.3.291] [PMID: 26082586]
[26]
Hamzaoui, K.; Houman, H.; Ben Dhifallah, I.; Kamoun, M.; Hamzaoui, A. Serum BAFF levels and skin mRNA expression in patients with Behçet’s disease. Clin. Exp. Rheumatol., 2008, 26(4)(Suppl. 50), S64-S71.
[PMID: 19026118]
[27]
Aksoy, Y.; Ercan, A.; Dalmizrak, O.; Canpinar, H.; Kartal Durmazlar, S.P.; Bayazit, M. The determination of matrix metalloproteinase 9 activity and gene expression levels in Behcet’s disease patients with aneurysmal complications. Clin. Rheumatol., 2011, 30(4), 515-519.
[http://dx.doi.org/10.1007/s10067-010-1559-3] [PMID: 20842516]
[28]
Ernst, M.; Inglese, M.; Scholz, G.M.; Harder, K.W.; Clay, F.J.; Bozinovski, S.; Waring, P.; Darwiche, R.; Kay, T.; Sly, P.; Collins, R.; Turner, D.; Hibbs, M.L.; Anderson, G.P.; Dunn, A.R. Constitutive activation of the SRC family kinase Hck results in spontaneous pulmonary inflammation and an enhanced innate immune response. J. Exp. Med., 2002, 196(5), 589-604.
[http://dx.doi.org/10.1084/jem.20020873] [PMID: 12208875]
[29]
Thomas, R.M.; Schmedt, C.; Novelli, M.; Choi, B.K.; Skok, J.; Tarakhovsky, A.; Roes, J. C-terminal SRC kinase controls acute inflammation and granulocyte adhesion. Immunity, 2004, 20(2), 181-191.
[http://dx.doi.org/10.1016/S1074-7613(04)00023-8] [PMID: 14975240]
[30]
Mazzi, P.; Caveggion, E.; Lapinet-Vera, J.A.; Lowell, C.A.; Berton, G. The Src-Family Kinases Hck and Fgr Regulate Early Lipopolysaccharide-Induced Myeloid Cell Recruitment into the Lung and Their Ability To Secrete Chemokines. J. Immunol., 2015, 195(5), 2383-2395.
[http://dx.doi.org/10.4049/jimmunol.1402011] [PMID: 26232427]
[31]
Fumagalli, L.; Campa, C.C.; Germena, G.; Lowell, C.A.; Hirsch, E.; Berton, G. Class I phosphoinositide-3-kinases and SRC kinases play a nonredundant role in regulation of adhesion-independent and -dependent neutrophil reactive oxygen species generation. J. Immunol., 2013, 190(7), 3648-3660.
[http://dx.doi.org/10.4049/jimmunol.1201951] [PMID: 23447687]
[32]
Lowell, C.A.; Berton, G. Resistance to endotoxic shock and reduced neutrophil migration in mice deficient for the Src-family kinases Hck and Fgr. Proc. Natl. Acad. Sci. USA, 1998, 95(13), 7580-7584.
[http://dx.doi.org/10.1073/pnas.95.13.7580] [PMID: 9636192]
[33]
Vicentini, L.; Mazzi, P.; Caveggion, E.; Continolo, S.; Fumagalli, L.; Lapinet-Vera, J.A.; Lowell, C.A.; Berton, G. Fgr deficiency results in defective eosinophil recruitment to the lung during allergic airway inflammation. J. Immunol., 2002, 168(12), 6446-6454.
[http://dx.doi.org/10.4049/jimmunol.168.12.6446] [PMID: 12055264]
[34]
Dallari, S.; Macal, M.; Loureiro, M.E.; Jo, Y.; Swanson, L.; Hesser, C.; Ghosh, P.; Zuniga, E.I. Src family kinases Fyn and Lyn are constitutively activated and mediate plasmacytoid dendritic cell responses. Nat. Commun., 2017, 8(14830), 14830.
[http://dx.doi.org/10.1038/ncomms14830] [PMID: 28368000]
[35]
Beavitt, S.J.; Harder, K.W.; Kemp, J.M.; Jones, J.; Quilici, C.; Casagranda, F.; Lam, E.; Turner, D.; Brennan, S.; Sly, P.D.; Tarlinton, D.M.; Anderson, G.P.; Hibbs, M.L. Lyn-deficient mice develop severe, persistent asthma: Lyn is a critical negative regulator of Th2 immunity. J. Immunol., 2005, 175(3), 1867-1875.
[http://dx.doi.org/10.4049/jimmunol.175.3.1867] [PMID: 16034130]
[36]
Marchetti, S.; Gamas, P.; Belhacène, N.; Grosso, S.; Pradelli, L.A.; Colosetti, P.; Johansen, C.; Iversen, L.; Deckert, M.; Luciano, F.; Hofman, P.; Ortonne, N.; Khemis, A.; Mari, B.; Ortonne, J.P.; Ricci, J.E.; Auberger, P. The caspase-cleaved form of LYN mediates a psoriasis-like inflammatory syndrome in mice. EMBO J., 2009, 28(16), 2449-2460.
[http://dx.doi.org/10.1038/emboj.2009.183] [PMID: 19590497]
[37]
Liossis, S.N.; Solomou, E.E.; Dimopoulos, M.A.; Panayiotidis, P.; Mavrikakis, M.M.; Sfikakis, P.P. B-cell kinase lyn deficiency in patients with systemic lupus erythematosus. J. Investig. Med., 2001, 49(2), 157-165.
[http://dx.doi.org/10.2310/6650.2001.34042] [PMID: 11288756]
[38]
Huck, S.; Le Corre, R.; Youinou, P.; Zouali, M. Expression of B cell receptor-associated signaling molecules in human lupus. Autoimmunity, 2001, 33(3), 213-224.
[http://dx.doi.org/10.3109/08916930109008048] [PMID: 11683380]
[39]
Bi, W.Y.; Fu, B.D.; Shen, H.Q.; Wei, Q.; Zhang, C.; Song, Z.; Qin, Q.Q.; Li, H.P.; Lv, S.; Wu, S.C.; Yi, P.F.; Wei, X.B. Sulfated derivative of 20(S)-ginsenoside Rh2 inhibits inflammatory cytokines through MAPKs and NF-kappa B pathways in LPS-induced RAW264.7 macrophages. Inflammation, 2012, 35(5), 1659-1668.
[http://dx.doi.org/10.1007/s10753-012-9482-1] [PMID: 22614119]
[40]
Guha, M.; Mackman, N. LPS induction of gene expression in human monocytes. Cell. Signal., 2001, 13(2), 85-94.
[http://dx.doi.org/10.1016/S0898-6568(00)00149-2] [PMID: 11257452]

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