Durán LM, Gaitán IR, Cano DL

Language: Spanish
References: 25
Page: 146-155
PDF size: 267.31 Kb.

ABSTRACT

Introduction: neutrophils are the first line of defense, releasing their granular contents made up of reactive oxygen species-generating enzymes such as myeloperoxidase and proteolytic enzymes such as elastase, which degrade elastin and collagen.
Objective: to evaluate toxicity of marine sponge Neopetrosia rosariensis extracts and their action as inhibitors of neutrophil degranulation and inhibition of the catalytic activity of human neutrophil elastase.
Method: the cytotoxicity was assessed by Trypan blue exclusion test of cell viability. Degranulation inhibition was evaluated by reduction of myeloperoxidase release percentage and inhibition of catalytic activity of elastase was determined by reduction of enzyme activity on specific substrate.
Results: the extracts exhibited low toxicity at the evaluated concentrations, with cellular viability percentage exceeding 80 % even at the highest concentration (25 µg/mL). They also showed inhibitory action of myeloperoxidase release, the greatest effect being concentrated on the most polar extracts (total and partial methanolic). The inhibition of the elastase activity was mostly found in the lowest polarity extracts (hexane and partial dichloromethane) at concentrations of 10 and 25 µg/mL, according to the hydrophobic character of the active site of the enzyme.
Conclusions: these results of the evaluated extracts are encouraging, considering each extract represents a complex mixture of active compounds of 10-25 µg/mL. This arouses great interest to continue their fractioning as potential agents for possible therapy of inflammatory processes.

REFERENCES

1. Lungarella G, Cavarra E, Lucattelli M, Martorana PA. The dual role of neutrophil elastase in lung destruction and repair. Intern J Biochem Cell Biol. 2008;40(6-7): 1287-96.

2. Kobayashi SD, Voyich JM, DeLeo FR. Regulation of the neutrophil-mediated inflammatory response to infection. Microbes and Infection. 2003;5(14):1337-44.

3. Vodovotz Y, Constantine G, Rubin J, Csete M, Voit EO, An G. Mechanistic simulations of inflammation: current state and future prospects. Mathematical Biosciences. 2009;217(1):1-10.

4. Le Y, Gong W, Tiffany HL, Tumanov A, Nedospasov S, Shen W, et al. Amyloid (beta) 42 activates a G-protein-coupled chemoattractant receptor, FPR-like-1. J Soc Neuroscience. 2001;21(2):RC123.

5. Selvatici R, Falzarano S, Mollica A, Spisani S. Signal transduction pathways triggered by selective formylpeptide analogues in human neutrophils. Eur J Pharmacol. 2006;534(1-3):1-11.

6. Le Y, Iribarren P, Zhou Y, Gong W, Hu J, Zhang X, et al. Silencing the formylpeptide receptor FPR by short-interfering RNA. Molecular Pharmacology. 2004;66(4):1022-8.

7. Bartolotta S, Scuteri M, Hick A, Palermo J, Rodriguez Brasco M, Hajdu E, et al. Evaluation of genotoxic biomarkers in extracts of marine sponges from Argentinean South Sea. J Exper Marine Biol Ecol. 2009;369(2):144-7.

8. Kernan M, Faulkner D, Parkanyi L, Clardy J, De Carvalho M, Jacobs R. Luffolide, a novel anti-inflammatory terpene from the spongeLuffariella sp. Cell Mol Life Sci. 1989;45(4):388-90.

9. Alcaraz MJ, Paya M. Marine sponge metabolites for the control of inflammatory diseases. Curr Opin Invest Drugs. 2006;7(11):974-9.

10. Franco LA, Macareno JL, Ocampo YC, Pájaro IB, Gaitán R. Marine sponges of the genus neopetrosia with anti-inflammatory activity. Latin Am J Pharm. 2012;31:976-83.

11. Strober W. Trypan blue exclusion test of cell viability. Curr Protocols Immunol. 2001:A. 3B. 1-A. 3B. 2.

12. Soto E, Limón A, Ortega A, Vega R. Características morfológicas y electrofisiológicas de las neuronas del ganglio vestibular en cultivo. Gac Méd Méx. 2002;138(1):1-13.

13. Johansson S, Göransson U, Luijendijk T, Backlund A, Claeson P, Bohlin L. A neutrophil multitarget functional bioassay to detect anti-inflammatory natural products. J Natural Products. 2002;65(1):32-41.

14. Edelson PJ, Cohn ZA. Peroxidase-mediated mammalian cell cytotoxicity. J Exper Med. 1973;138(1):318-23.

15. Henson PM, Zanolari B, Schwartzman NA, Hong SR. Intracellular control of human neutrophil secretion. J Immunol. 1978;121(3):851-5.

16. Siedle B, Gustavsson L, Johansson S, Murillo R, Castro V, Bohlin L, et al. The effect of sesquiterpene lactones on the release of human neutrophil elastase. Biochemical Pharmacol. 2003;65(5):897-903.

17. Kanashiro A, Souza JG, Kabeya LM, Azzolini A, Lucisano-Valim YM. Elastase release by stimulated neutrophils inhibited by flavonoids: importance of the catechol group. Zeitschrift fur Naturforschung C-Journal of Biosciences. 2007;62(5-6): 357-61.

18. Lentini A, Ternai B, Ghosh P. Synthetic inhibitors of human granulocyte elastase, Part 4. Inhibition of human granulocyte elastase and cathepsin G by nonsteroidal anti-inflammatory drugs (NSAID) s. Biochem Inter. 1987;15(6):1069.

19. Kang K, Bae SJ, Kim WM, Lee DH, Cho U, Lee MH, et al. Molecular characteristics of the inhibition of human neutrophil elastase by nonsteroidal antiinflammatory drugs. Exper Mol Med. 2000;32(3):146.

20. Escrig V, Ubeda A, Ferrandiz M, Darias J, Sanchez J, Alcaraz M, et al. Variabilin: a dual inhibitor of human secretory and cytosolic phospholipase A2 with antiinflammatory activity. J Pharmacol Exper Therap. 1997;282(1):123-31.

21. Johansson S. Studies on cytotoxic and neutrophil challenging polypeptides and cardiac glycosides of plant origin. Uppsala University; 2001.

22. Keyzers RA. The isolation of biologically active secondary metabolites from New Zealand marine organisms [[PhD Thesis]. New Zealand: Victoria University of Wellington; 2003. p. 163-7.

23. Keyzers RA, Davies-Coleman MT. Anti-inflammatory metabolites from marine sponges. Chem Soc Rev. 2005;34(4):355-65.

24. Hong S, Kim SH, Rhee MH, Kim AR, Jung JH, Chun T, et al. In vitro anti-inflammatory and pro-aggregative effects of a lipid compound, petrocortyne A, from marine sponges. Naunyn-Schmiedeberg's Arch Pharmacol. 2003;368(6):448-56.

25. Shapiro S. Proteinases in chronic obstructive pulmonary disease. Biochemical Society Transact. 2002;30:98-102.