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A fucan of a brown seaweed and its antitumoral property on HT-29 and immunomodulatory activity in murine RAW 264.7 macrophage cell line

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Abstract

Polysaccharides from algae are also proper candidates with therapeutic properties and immunomodulatory and antitumor effects. The brown seaweed Lobophora variegata synthesizes different groups of anionic polysaccharides with several biological properties. Sulfated polysaccharides were obtained by delipidation of seaweed, proteolysis, and fractionation with different volumes of acetone. A fraction of sulfated polysaccharides, fucans or fucoidans, was extracted with 0.8 v acetone and named L. variegata (LV). This fucan was assessed in the inflammatory process in rats, antitumor action on human colon adenocarcinoma (HT-29 cells), apoptosis, and its effect on the cell cycle. LV fraction, a galactofucan, exhibited a high ratio of total sugar/sulfate (1.5) and a very low level of proteins. This polysaccharide showed an antiinflammatory effect on two models of inflammation induced by croton oil and oxazolone in rats. LV was analyzed in cellular proliferation of HT-29. We also demonstrated the cytotoxic action against this cell line and induction in the apoptosis and decreased the cell cycle in phases S and G 2/M and the accumulation of cells in the G 1 phase. Our studies with LV showed a marked immunomodulatory action without and with lipopolysaccharide (LPS) on RAW 264.7 cells. Comparative studies of two fucans, LV and FV (Fucus vesiculosus), with different structures were assessed on viability in macrophages, RAW 264.7 cells. Both showed cytotoxicity in these cells. We observed high levels of nitric oxide (NO) production, interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) when treated at 0.25, 0.5, and 1.0 mg mL−1 of LV. Data also suggest that LV has potential antitumor effects on HT-29 and antiinflammatory and immunomodulatory effects on RAW 264.7 cells.

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References

  • Aisa Y, Miyakawa Y, Nakazato T, Shibata H, Saito K, Ikeda Y, Kizaki M (2005) Fucoidan induces apoptosis of human HS-sultan cells accompanied by activation of caspase-3 and down-regulation of ERK. Am J Hematol 78:7–14

    Article  CAS  PubMed  Google Scholar 

  • Ale MT, Mikkelsen JD, Meyer AS (2011) Important determinants for fucoidan bioactivity: a critical review of structure-function relations and extraction methods for fucose-containing sulfated polysaccharides from brown seaweeds. Mar Drugs 9:2106–2130

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Alves MGCF, Dore CMPG, Castro AJG, Nascimento MS, Cruz AKM, Soriano ME, Benevides MB, Leite EL (2012) Antioxidant, cytotoxic and hemolytic effects of sulfated galactans from edible red alga Hypnea musciformis. J Appl Phycol 24:1217–1227

    Article  CAS  Google Scholar 

  • Azevedo TCG, Bezerra MEB, Santos MGL, Souza LA, Marques CT, Benevides NMB, Leite EL (2009) Heparinoids algal and their anticoagulant, hemorrhagic activities and platelet aggregation. Biom Pharm 63:477–483

    Article  Google Scholar 

  • Beekes M, Lasch P, Naumann D (2007) Analytical applications of Fourier transform-infrared (FT-IR) spectroscopy in microbiology and prion research. Vet Microbiol 123:305–319

    Article  CAS  PubMed  Google Scholar 

  • Berteau O, Mulloy B (2003) Sulfated fucans, fresh perspectives: structures, functions, and biological properties of sulfated fucans and an overview of enzymes active toward this class of polysaccharide. Glycobiology 13:29–40

    Article  Google Scholar 

  • Bilan MI, Grachev AA, Ustuzhanina NE, Shashkov AS, Nifantiev NE, Usov AI (2004) A highly regular fraction of a fucoidan from the brown seaweed Fucus distichus L. Carbohyd Res 339:511–517

    Article  CAS  Google Scholar 

  • Bilan MI, Grachev AA, Shashkov AS, Nifantiev NE, Usov A (2006) I. Structure of a fucoidan from the brown seaweed Fucus serratus. Carbohydr Res 341:238–245

    Article  CAS  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Carvalho AC, Sousa RB, Franco AX, Costa JV, Neves LM, Ribeiro RA, Sutton R, Criddle DN, Soares PM, De Souza MH (2014) Protective effects of fucoidan, a P- and L-selectin inhibitor, in murine acute pancreatitis. Pancreas 43:82–87

    Article  CAS  PubMed  Google Scholar 

  • Castro LSEPW, Pinheiro TS, Castro AJG, Dore CMPG, Silva NB, Alves MGCF, Santos MSN, Leite EL (2014) Fucose-containing sulfated polysaccharides from brown macroalgae Lobophora variegata with antioxidant, anti-inflammatory, and antitumoral effects. J Appl Phycol 26:1783–1790

    Article  CAS  Google Scholar 

  • Castro LSEPW, Pinheiro TS, Castro AJG, Santos MSN, Soriano EM, Leite EL (2015) Potential anti-angiogenic, antiproliferative, antioxidant and anticoagulant activity of anionic polysaccharides, fucans, extracted from brown algae Lobophora variegata. J Appl Phycol 27:1315–1325

    Article  CAS  Google Scholar 

  • Castro LSEPW, Castro AJG, Pinheiro TS, Santos MSN, Alves LG, Soriano EM, Araújo RM, Leite EL (2016) Effect of galactofucan of a brown seaweed on induced hepatoxicity in rats, sodium pentobarbital-induced sleep, and anti-inflammatory activity. J Appl Phycol 28:2005–2017

    Article  CAS  Google Scholar 

  • Chapman ALP, Mocatta TJ, Shiva S, Seidel A, Chen B, Khalilova I, Paumann-Page ME, Jameson GNL, Winterbourn CC, Kettle AJ (2013) Ceruloplasmin is an endogenous inhibitor of myeloperoxidase. J Biol Chem 288:6465–6477

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cho M, Lee D, Kim J, You S (2014) Molecular characterization and immunomodulatory activity of sulfated fucans from Agarum cribrosum. Carbohydr Polym 113:507–514

    Article  CAS  PubMed  Google Scholar 

  • Coleman JW (2001) Nitric oxide in immunity and inflammation. Int Immunopharmacol 1:1397–1406

    Article  CAS  PubMed  Google Scholar 

  • Commins SP, Borish L, Steinke JW (2010) Immunologic messenger molecules: cytokines, interferons, and chemokines. J Allergy Clin Immunol 125:53–72

    Article  Google Scholar 

  • Croci DO, Cumashi A, Ushakova NA, Preobrazhenskaya ME, Piccoli A, Totani L, Ustyuzhanina NE, Bilan MI, Usov AI, Grachev AA, Morozevich GE, Berman AE, Sanderson CJ, Kelly M, Di Gregorio P, Rossi C, Tinari N, Iacobelli S, Rabinovich GA, Nifantiev NE (2011) Fucans, but not fucomannoglucuronans, determine the biological activities of sulfated polysaccharides from Laminaria saccharina brown seaweed. PLoS One 6:e17283

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cumashi A, Ushakova NA, Preobrazhenskaya ME, D'Incecco A, Piccoli A, Totani L, Tinari N, Morozevich GE, Berman AE, Bilan MI, Usov AI, Ustyuzhanina NE, Grachev AA, Sanderson CJ, Kelly M, Rabinovich GA, Iacobelli S, Nifantiev NE (2007) A comparative study of the anti-inflammatory, anticoagulant, antiangiogenic, and antiadhesive activities of nine different fucoidans from brown seaweeds. Glycobiology 17:541–552

    Article  CAS  PubMed  Google Scholar 

  • Damonte EB, Matulewicz MC, Cerezo AS (2004) Sulfated seaweed polysaccharides as antiviral agents. Curr Med Chem 11:2399–2419

    Article  CAS  PubMed  Google Scholar 

  • De Young LM, Kheifets JB, Ballaron SJ, Young JM (1989) Edema and cell infiltration in the phorbol ester-treated mouse ear are temporally separated and can be differentially modulated by pharmacologic agents. Agents Actions 26:335–341

    Article  CAS  PubMed  Google Scholar 

  • Dhillon AS, Hagan S, Rath O, Kolch W (2007) MAP kinase signalling pathways in cancer. Oncogene 26:3279–3290

    Article  CAS  PubMed  Google Scholar 

  • Dodgson KS, Price RG (1962) A note on the determination of the ester sulphate content of sulphated polysaccharides. Biochem J 84:106–110

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dore CMPG, Alves MGCF, Santos ND, Cruz AKM, Camara RBG, Castro AJG, Alves LG, Nader HB, Leite EL (2013) Antiangiogenic activity and direct antitumor effect from a sulfated polysaccharide isolated from seaweed. Microvasc Res 88:12–18

    Article  Google Scholar 

  • Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars, and related substances. Anal Chem 28:350–356

    Article  CAS  Google Scholar 

  • Duque GA, Desconteaux A (2014) Macrophage cytokines: involvement in immunity and infectious diseases. Front Immunol 5:491. doi:10.3389/fimmu.2014.00491

    Google Scholar 

  • Ferreri NR, Millet I, Paliwal V, Herzog W, Solomon D, Ramabhadran R, Askenase PW (1991) Induction of macrophage TNF alpha, IL-1, IL-6, and PGE2 production by DTH-initiating factors. Cell Immunol 137:389–405

    Article  CAS  PubMed  Google Scholar 

  • Foster I (2008) Cancer: a cell cycle defect. Radiography 14:144–149

    Article  Google Scholar 

  • Gábor M (2000) Mouse ear inflammation models and their pharmacological applications. Akadémiai Kiadó, Budapest

    Google Scholar 

  • Gomez-Ordonez E, Ruperez P (2011) FTIR-ATR spectroscopy as a tool for polysaccharide identification in edible brown and red seaweeds. Food Hydrocoll 25:1514–1520

    Article  CAS  Google Scholar 

  • Hajiaghaalipour F, Kanthimathi MS, Sanusi J, Rajarajeswaran J (2015) White tea (Camellia sinensis) inhibits proliferation of the colon cancer cell line, HT-29, activates caspases and protects DNA of normal cells against oxidative damage. Food Chem 169:401–410

    Article  CAS  PubMed  Google Scholar 

  • Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144:646–674

    Article  CAS  PubMed  Google Scholar 

  • Hunziker T, Kolma RA, Spath PJ, Braathen LR (1992) Circulating immune complexes in patients with psoriasis: do they exist? Exp Dermatol 3:149–151

    Article  Google Scholar 

  • Ignaro LJ (1996) Nitric oxide as a communication signal in vascular and neuronal cells. In: Lancaster J (ed). Nitric oxide—principles and actions. New York: Academic Press pp 111–35

  • Jin JO, Yu Q (2015) Fucoidan delays apoptosis and induces pro-inflammatory cytokine production in human neutrophils. Int J Biol Macromol 73:65–71

    Article  CAS  PubMed  Google Scholar 

  • Juan ME, Wenzel U, Daniel H, Planas JM (2008) Resveratol induces apoptosis through ROS-dependent mitochondria pathway in HT-29 human colorectal carcinoma cell. J Agric Food Chem 56:4813–4818

    Article  CAS  PubMed  Google Scholar 

  • Jung CH, Kim JH, Park SJ, Kweon DH, Kim SH, Ko S-G (2010) Inhibitory effect of Agrimonia pilosa Ledeb on inflammation by suppression of iNOS and ROS production. Immunol Invest 39:159–170

  • Karmakar P, Ghosh T, Sinha S, Saha S, Mandal P, Ghosal PK, Ray B (2009) Polysaccharides from the brown seaweed Padina tetrastromatica: characterization of a sulfated fucan. Carbohyd Polym 78:416–421

    Article  CAS  Google Scholar 

  • Karpuzoglu E, Ahmed SA (2006) Estrogen regulation of nitric oxide and inducible nitric oxide synthase (iNOS) in immune cells: implications for immunity, autoimmune diseases, and apoptosis. Nitric Oxide J 15:177–186

    Article  CAS  Google Scholar 

  • Kettle AJ, Winterbourn CC (1991) Mechanism of inhibition of myeloperoxidase by anti-inflammatory drugs. Biochem Pharmacol 41:1485–1492

    Article  CAS  PubMed  Google Scholar 

  • Leite ED, Medeiros MGL, Rocha HAO, Farias GGM, Silva LF, Chavante SF, LRD A, Dietrich CP, Nader HB (1998) Structure and pharmacological activities of sulfated xylofucoglucuronan from the alga Spatoglossum schröederi. Plant Sci 132:215–228

    Article  CAS  Google Scholar 

  • Li Y, Huang W, Huang S, Du J, Huang JC (2012) Screening of anti-cancer agent using zebrafish: Comparison with the MTT assay. Biochem Biophys Res Commun 422:85–90

  • Medeiros VP, Queiroz KCS, Cardoso ML, Monteiro GRG, Oliveira FW, Chavante SF, Guimarães LA, Rocha HAO, Leite EL (2008) Sulfated galactofucan from Lobophora variegata: anticoagulant and anti-inflammatory proprieties. Biochemistry 73:1265–1273

    Google Scholar 

  • Moncada M, Palmer RMJ, Higges EA (1991) Nitric oxide: physiology, pathophysiology and pharmacology. Pharmacol Rev 43:109–142

    CAS  PubMed  Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    Article  CAS  PubMed  Google Scholar 

  • Murphy M, Hotchkiss S, Worthington J, McKeown SR (2014) The potential of seaweed as a source of drugs for use in cancer chemotherapy. J Appl Phycol 26:2211–2264

    Article  CAS  Google Scholar 

  • Nasu T, Fukuda Y, Nagahira K, Kawashima H, Noguchi C, Nakanishi T (1997) Fucoidin, a potent inhibitor of L-selectin function, reduces contact hypersensitivity reaction in mice. Immunol Lett 59:47–51

    Article  CAS  PubMed  Google Scholar 

  • Nishino T, Nagumo T (1992) Anticoagulant and antithrombin activities of oversulfated fucans. Carbohydr Res 229:355–362

    Article  CAS  PubMed  Google Scholar 

  • Nobre LTDB, Vidal AAJ, Almeida-Lima J, Oliveira RM, Paredes-Gameiro EJ, Medeiros VP, Trindade EF, Franco CRC, Nader HB, Rocha HAO (2013) Fucan effect on CHO cell proliferation and migration. Carbohydr Polym 98:224–232

    Article  CAS  PubMed  Google Scholar 

  • Nousbaum A, Vocanson M, Rozieres A, Hennino A, Nicolas JF (2009) Allergic and irritant contact dermatitis. Eur J Dermatol 19:325–332

    Google Scholar 

  • Oda T, Maeda H (1986) A new simple fluorometric assay for phagocytosis. J Immunol Meth 88:175–183

    Article  CAS  Google Scholar 

  • Paiva AAO, Castro AJG, Nascimento MS, Will LSEP, Santos ND, Araújo RM, Xavier CAC, Rocha FA, Leite EL (2011) Antioxidant and antiinflammatory effect of polysaccharides from Lobophora variegata on zymosan-induced arthritis in rats. Int Immunopharm 11:1241–1250

    Article  CAS  Google Scholar 

  • Podrez E, Abu-Soud HM, Hazen SL (2000) Myeloperoxidase-generated oxidants and atherosclerosis. Free Radic Biol Med 28:1717–1725

    Article  CAS  PubMed  Google Scholar 

  • Prieto JM, Recio MC, Giner RM (2006) Anti-inflammatory activity of β-sitosterol in a model of oxazolone induced contact-delayed-type hypersensitivity. Bol Latinoam Caribe Plant Med Arom 5:57–62

    Google Scholar 

  • Reagan-Shaw S, Nihal M, Ahmad N (2007) Dose translation from animal to human studies revisited. FASEB J 22:659–661

    Article  PubMed  Google Scholar 

  • Recio MC, Giner RM, Uriburu L, Máñez S, Cerdá M, De La Fuente JR, Rios JL (2000) In vivo activity of pseudoguaianolide sesquiterpene lactones in acute and chronic inflammation. Life Sci 66:2509–2518

    Article  CAS  PubMed  Google Scholar 

  • Rocha HAO, Franco CRC, Trindade ES, Carvalho LCM, Veiga SS, Leite EL, Dietrich CP, Nader HB (2001) A fucan from the brown seaweed Spatoglossum schröederi inhibits Chinese hamster ovary cell adhesion to several extracellular matrix proteins. Braz Med Biol Res 34:621–626

    Article  CAS  Google Scholar 

  • Rocha HAO, Moraes FA, Trindade ES, Franco CRC, Torquato RJC, Velga SS, Valente AP, Mourão PAS, Leite EL, Nader HB, Dietrich CP (2005) Structural and hemostatic activities of a sulfated galactofucan from the brown alga Spatoglossum schröederi an ideal antithrombotic agent? J Biol Chem 280:41278–41288

    Article  CAS  PubMed  Google Scholar 

  • Rossol M, Heine H, Meusch U, Quandt D, Klein C, Sweet MJ, Hauschildt S (2011) LPS-induced cytokine production in human monocytes and macrophages. Crit Rev Immunol 31:379–446

    Article  CAS  PubMed  Google Scholar 

  • Salatino A, Salatino MLF, Negri G (2007) Traditional uses, chemistry and pharmacology of Croton species (Euphorbiaceae). J Braz Chem Soc 18:11–33

    Article  CAS  Google Scholar 

  • Schoepe S, Schäcke H, May E, Asadullah K (2006) Glucocorticoid therapy-induced skin atrophy. Exp Dermatol 15:406–420

    Article  CAS  PubMed  Google Scholar 

  • Senthilkumar K, Manivasagan P, Venkatesan J, Kim S-K (2013) Brown seaweed fucoidan: biological activity and apoptosis, growth signaling mechanism in cancer. Int J Biol Macromol 60:366–374

    Article  CAS  PubMed  Google Scholar 

  • Souza MCR, Marques CT, Dore CMPG, Silva FRF, Rocha HAO, Leite EL (2007) Antioxidant activities of sulfated polysaccharides from brown and red seaweeds. J Appl Phycol 19:153–160

    Article  Google Scholar 

  • Suzuki K, Ota H, Sasagawa S, Sakatani T, Fujikura T (1983) Assay method for myeloperoxidase in human polymorphonuclear leukocytes. Anal Biochem 132:345–352

    Article  CAS  PubMed  Google Scholar 

  • Tedder TF, Steeber DA, Chen A, Engel P (1995) The selectins: vascular adhesion molecules. FASEB J 9:866–873

    CAS  PubMed  Google Scholar 

  • Tubaro A, Dri P, Delbello G, Zilli C, Della Loggia R (1985) The cróton oil ear test revisited. Agents Actions 17:347–349

    Article  CAS  Google Scholar 

  • Tuschl H, Schwab CE (2004) Flow cytometric methods used as screening tests for basal toxicity of chemicals. Toxicol in Vitro 18:483–491

    Article  CAS  PubMed  Google Scholar 

  • Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Talser J (2007) Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell B 39:44–84

    Article  CAS  Google Scholar 

  • Wlodkowic D, Telford W, Skommer J, Darzynkiewicz Z (2011) Apoptosis and beyond: cytometry in studies of programmed cell death. Methods Cell Biol 103:55–98

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zemelka-Wiacek M, Majewska-Szczepanik M, Pyrczak W, Szczepanik M (2013) Complementary methods for contact hypersensitivity (CHS) evaluation in mice. J Immunol Meth 387:270–275

    Article  CAS  Google Scholar 

  • Zhang R, Brennan ML, Shen Z, MacPherson JC, Schmitt D, Molenda CE, Hazen SL (2002) Myeloperoxidase functions as a major enzymatic catalyst for initiation of lipid peroxidation at sites of inflammation. J Biol Chem 277:46116–46122

    Article  CAS  PubMed  Google Scholar 

  • Zhang W, Du JY, Jiang ZD, Okimura T, Oda T, Yu Q, Jin J-O (2014) Ascophyllan purified from Ascophyllum nodosum induces Th1 and Tc1 immune responses by promoting dendritic cell maturation. Mar Drugs 12:4148–4164

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhao J, Bolton EM, Bradley JA, Lever AML (2009) Lentiviral-mediated overexpression of Bcl-xL protects primary endothelial cells from ischemia/reperfusion injury-induced apoptosis. J Heart Lung Transplant 28:936–943

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  1. Laboratory of Glycobiology, Department of Biochemistry, Federal University of Rio Grande do Norte, Av. Salgado Filho, 3000, Bairro L. Nova, CEP, Natal, RN, 59072-970, Brazil

    Thuane de Sousa Pinheiro, Marilia da S. Nascimento Santos, Luiza Sheyla E. P. Will Castro, Almino Afonso de O. Paiva, Luciana G. Alves, Ana Katarina M. Cruz, Leonardo Thiago D. B. Nobre, Monique Gabriela das C. F. Alves & Edda Lisboa Leite

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  1. Thuane de Sousa Pinheiro
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  2. Marilia da S. Nascimento Santos
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  3. Luiza Sheyla E. P. Will Castro
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  4. Almino Afonso de O. Paiva
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  5. Luciana G. Alves
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  6. Ana Katarina M. Cruz
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de Sousa Pinheiro, T., Nascimento Santos, M.d.S., Will Castro, L.S.E.P. et al. A fucan of a brown seaweed and its antitumoral property on HT-29 and immunomodulatory activity in murine RAW 264.7 macrophage cell line. J Appl Phycol 29, 2061–2075 (2017). https://doi.org/10.1007/s10811-017-1075-9

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