Preliminary molecular characterization of a proinflammatory and nociceptive molecule from the Echinometra lucunter spines extracts

Juliana Mozer Sciani1, Bianca Zychar2, Luis Roberto Gonçalves2, Renata Giorgi2, Thiago Nogueira2, Daniel Carvalho Pimenta1
1Laboratory of Biochemistry and Biophysics, Butantan Institute, Butantã, Brazil
2Laboratory of Pathophysiology, Butantan Institute, Butantã, Brazil

Tóm tắt

Sea urchins are animals commonly found on the Brazilian shoreline, being Echinometra lucunter the most abundant species. Accidents caused by E. lucunter have been reported as one of the most frequent in Brazil, and are characterized by intense pain and inflammation, consequence of spine puncture in the skin. In order to characterize such toxic effects, we isolated one molecule that caused inflammatory and nociceptive effects. E. lucunter specimens were collected without gender distinction. Spines were removed and molecules were extracted, fractionated by RP-HPLC and assayed for inflammatory and nociceptive activity, in a biological-driven fractionation way, until the obtainment of one active molecule and its subsequent analysis by mass spectrometry (MS and MS/MS). For inflammation, intravital microscopy was performed on the mouse cremaster muscle, in order to evaluate rolled, adherent and migrating leukocytes. Paw edema was also evaluated. For the nociceptive activity, the paw pressure test was performed in rats. One molecule could be isolated and related to the inflammatory and nociceptive activity. Regarding inflammation, increase in adherent and migrating cells was observed in the cremaster muscle after the administration of the molecule. Corroborating the inflammatory response, paw edema was also observed, although only in 20% of controls and 20 min after injection. Additionally, this molecule was able to decrease significantly the pain threshold, characterizing hyperalgesia. This molecule was analyzed by mass spectrometry, and according to the exact molecular mass, isotopic distribution and fragmentation profile, it was possible to propose the molecular formula C29H48N3O10. One isolated molecule from the spine extract of E. lucunter is able to elicit inflammation and hypernociception in animal models, which is in agreement with the effects observed in sea urchin accidents.

Tài liệu tham khảo

Hiratsuka Y, Uehara T. Feeding rates and absorption efficiencies of four species of sea urchins (genus Echinometra) fed a prepared diet. Comp Biochem Physiol A Mol Integr Physiol. 2007;148(1):223–9. McPherson BF. Studies on the biology of the tropical sea urchins, Echinometra lucunter and Echinometra viridis. Bull Mar Sci. 1969;19(1):194–213. Haddad V Jr, Lupi O, Lonza JP, Tyring SK. Tropical dermatology: marine and aquatic dermatology. J Am Acad Dermatol. 2009;61(5):733–50. Smith ML. Skin problems from marine echinoderms. Dermatol Ther. 2002;15:30–3. Nassab R, Rayatt S, Peart F. The management of hand injuries caused by sea urchin spines. J Hand Surg Br. 2005;30(4):432–3. Rocha G, Fraga S. Sea urchin granuloma of the skin. Arch Dermatol. 1962;85:406–8. Kabigting FD, Kempiak SJ, Alexandrescu DT, Yu BD. Sea urchin granuloma secondary to Strongylocentrotus purpuratus and Strongylocentrotus franciscanus. Dermatol Online J. 2009;15(5):9. Sciani JM, Antoniazzi MM, Neves AC, Pimenta DC. Cathepsin B/X is secreted by Echinometra lucunter sea urchin spines, a structure rich in granular cells and toxins. J Venom Anim Toxins incl Trop Dis. 2013;19:33. doi:10.1186/1678-9199-19-33. Sciani JM, Zychar BC, Gonçalves LR, Nogueira Tde O, Giorgi R, Pimenta DC. Pro-inflammatory effects of the aqueous extract of Echinometra lucunter sea urchin spines. Exp Biol Med (Maywood). 2011;236(3):277–80. Sciani JM, Emerenciano AK, Cunha da Silva JR, Pimenta DC. Initial peptidomic profiling of Brazilian sea urchins: Arbacia lixula, Lytechinus variegatus and Echinometra lucunter. J Venom Anim Toxins incl Trop Dis. 2016;22:17. doi:10.1186/s40409-016-0071-x. Baez S. An open cremaster muscle preparation for the study of blood vessels by in vivo microscopy. Microvasc Res. 1973;5(3):384–94. Randall LO, Selitto JJ. A method for measurement of analgesic activity on inflamed tissue. Arch Int Pharmacodyn Ther. 1957;111(4):409–19. Rossetto AL, Mora JM, Haddad V Jr. Sea urchin granuloma. Rev Inst Med Trop S Paulo. 2006;48(5):303–6. Oldham ED, Seelam S, Lema C, Aguilera RJ, Fiegel J, Rankin SE, et al. Synthesis, surface properties and biocompatibility of 1,2,3-triazole-containing alkyl β-D-xylopyranoside surfactants. Carbohydr Res 2013;379:68-77. Oldham ED, Nunes LM, Varela-Ramirez A, Rankin SE, Knutson BL, Aguilera RJ, et al. Cytotoxic activity of triazole-containing alkyl β-D-glucopyranosides on a human T-cell leukemia cell line. Chem Cent J. 2015;9:3. Blunt JW, Copp BR, Munro MHG, Norhcote PT, Prinsep MR. Marine natural products. Nat Prod Rep. 2003;20(1):1–48. Blunt JW, Copp BR, Keyzers RA, Munro MHG, Prinsep MR. Marine natural products. Nat Prod Rep. 2016;33(3):382–431. Blunt JW, Copp BR, Hu WP, Munro MHG, Northcote PT, Prinsep MR. Marine natural products. Nat Prod Rep. 2007;24(1):31–86. Binyon J. Physiology of echinoderms: international series of monographs in pure and applied biology zoology division. Paperback. Oxford: Pergamon; 2013. Mischenko NP, Fedoreyev SA, Pokhilo ND, Anufriev VP, Denisenko VA, Glazunov VP. Echinamines A and B, first aminated hydroxynaphthazarins from the sea urchin Scaphechinus mirabilis. J Nat Prod. 2005;68(9):1390–3. Kita M, Watanabe M, Takada N, Suenaga K, Yamada K, Uemura D. Hedathiosulfonic acids A and B, novel thiosulfonic acids from the deep-sea urchin Echinocardium cordatum. Tetrahedron. 2002;58:6405–12. Blunt JW, Munro MHG. Dictionary of marine natural products. Boca Raton: Taylor & Francis Group. 2008; Doi:10.1002/anie.200785567. Granger DN, Kubes P. The microcirculation and inflammation: modulation of leukocyte-endothelial cell adhesion. J Leukoc Biol. 1994;55(5):662–75.