Anti-Inflammatory and Apoptotic Activities of Pomolic Acid Isolated from Cecropia pachystachya

Abstract

The dichloromethane extract and pomolic acid (5) obtained from leaves of Cecropia pachystachya both reduced carrageenan-induced paw oedema in mice. Interestingly, while the triterpenoid inhibited the in vivo production of interleukin-1β by 39 %, it had no effect on tumour necrosis factor-α production. We also demonstrated that both the dichloromethane extract and 5 inhibited the viability of human polymorphonuclear (PMN) cells in a time- and dose-dependent fashion. The PMN membrane integrity was determined with the aid of flow cytometry by means of the exclusion of propidium iodide as assay. Although the cell membrane integrity was altered, neither the extract nor 5 produced cellular necrosis. Moreover, the development of hypodiploid nuclei and DNA fragmentation in the PMN cells were both dependent on dose and time. Finally, in the annexin V-FITC binding assay, compound 5 increased the total of apoptotic cells by 42 % at 100 μM and by 71 % at 200 μM with respect to the control group. In conclusion, both the dichloromethane extract of ambay and isolated compound 5 inhibit the viability of PMN cells through apoptosis. Since they can regulate human neutrophil functions in this way, it is likely that these substances can also limit inflammation.

Abbreviations

DMSO:dimethyl sulfoxide

FITC:fluorescein isothiocyanate

IL-1β:interleukin-1β

LPS:lipopolysaccharide

MTT:3-(4,5-dimethylthiazolyl)-2,5-diphe-nyltetrazolium bromide

PBS:phosphate-buffered saline

PMN:polymorphonuclear

TNF-α:tumour necrosis factor-α

Ambay - Cecropia pachystachya - Moraceae - pomolic acid - anti-inflammatory activity - apoptosis - neutrophils

References

• 1 Consolini A E, Migliori G N. Cardiovascular effects of the South American medicinal plant Cecropia pachystachya (ambay) on rats.  J Ethnopharmacol. 2005;  96 417-22
• 2 Consolini A E, Ragone M I, Migliori G N, Conforti P, Volonté M G. Cardiotonic and sedative effects of Cecropia pachystachya Mart. (ambay) on isolated rat hearts and conscious mice.  J Ethnopharmacol. 2006;  106 90-6
• 3 Soraru S B, Bandoni A L. Plantas de la medicina popular argentina. Buenos Aires; Editorial Albatros 1978: 76-8
• 4 Petenatti E M, Petenatti M E, Del Vitto L A. Herbal remedies in Central-Western Argentina. ”Ambay”. Quality control of the genuine drug and their adulterants.  Acta Farm Bonaerense. 1998;  17 197-212
• 5 Juan-Hikawczuk V, Saad J R, Guardia T, Juarez A O, Giordano O S. Anti-inflammatory activity of compounds isolated from Cecropia pachystachya.  An Asoc Quim Arg. 1998;  86 167-70
• 6 Giner R M, Villalba M L, Recio M C, Máñez S, Cerdá-Nicolás M, Ríos J L. Anti-inflammatory glycoterpenoids from Scrophularia auriculata.  Eur J Pharmacol. 2000;  389 243-52
• 7 Escandell J M, Recio M C, Giner R M, Máñez S, Cerdá-Nicolás M, Ríos J L. Cucurbitacin R reduces the inflammation and bone damage associated with adjuvant arthritis in Lewis rats by suppression of TNF-α in T-lymphocytes and macrophages.  J Pharmacol Exp Ther. 2007;  320 581-90
• 8 Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.  J Immunol Methods. 1983;  65 55-63
• 9 Cruz-Chamorro L, Puertollano M A, Puertollano E, Álvarez de Cienfuegos G, de Pablo M A. In vitro biological activities of magainin alone or in combination with nisin.  Peptides. 2006;  27 1201-9
• 10 Nicoletti I, Megliorati G, Pagliacci M C, Grignani F, Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry.  J Immunol Methods. 1991;  139 271-9
• 11 Breitmaier E. Structure elucidation by NMR in organic chemistry. A practical guide. Chichester; Wiley 1993: 56
• 12 Brown G D. Phytene-1,2-diol from Artemisia annua.  Phytochemistry. 1994;  36 1553-4
• 13 Mahato S B, Kundu A P. ¹³C-NMR spectra of pentacyclic triterpenoids. A compilation and some salient features.  Phytochemistry. 1994;  37 1517-75
• 14 Prieto J M. Efecto de plantas antiinflamatorias en la función leucocitaria y procesos biooxidativos. Valencia (España); University of Valencia 2001: 117-9
• 15 Huguet A I, Recio M C, Máñez S, Giner R M, Ríos J L. Effect of triterpenoids on the inflammation induced by protein kinase C activators, neuronally-acting irritants and other agents.  Eur J Pharmacol. 2000;  410 69-81
• 16 Máñez S, Recio M C, Giner R M, Ríos J L. Effect of selected triterpenoids on chronic dermal inflammation.  Eur J Pharmacol. 1997;  334 103-5
• 17 Ríos J L, Recio M C, Máñez S, Giner R M. Natural triterpenoids as anti-inflammatory agents. In: Atta-Ur-Rahman, editor Studies in natural products chemistry: bioactive natural products, Vol. 22. Amsterdam; Elsevier Science Publishers 2000: 93-143
• 18 Rojano B, Pérez E, Figadère B, Martin M T, Recio M C, Giner R. et al . Constituents of Oxandra cf. xylopioides with anti-inflammatory activity.  J Nat Prod. 2007;  70 835-8
• 19 Giner E M, Máñez S, Recio M C, Giner R M, Cerdá M, Ríos J L. In vivo studies on the anti-inflammatory activity of pachymic and dehydrotumulosic acids.  Planta Med. 2000;  66 221-7
• 20 Escandell J M, Recio M C, Giner R M, Máñez S, Gil R, López-Ginés C. et al . Dihydrocucurbitacin B inhibits delayed-type hypersensitivity reaction by suppressing lymphocyte proliferation.  J Pharmacol Exp Ther. 2007;  322 1261-8
• 21 Savill J. Apoptosis in resolution of inflammation.  Kidney Blood Press Res. 2000;  23 173-4
• 22 Smith J A. Neutrophils, host defense and inflammation: a double edged sword.  J Leukoc Biol. 1994;  56 672-86
• 23 Chilvers E R, Cadwallader K A, Reed B J, White J F, Condliffe A M. The function and fate of neutrophils at the inflamed site: prospects for therapeutic intervention.  J R Coll Physicians Lond. 2000;  34 68-74
• 24 Downey G P, Fukushima T, Fialkow L, Waddell T K. Intracellular signaling in neutrophil priming and activation.  Semin Cell Biol. 1995;  6 345-56
• 25 Ríos J L, Recio M C. Natural products as modulators of apoptosis and their role in inflammation. In: Atta-Ur-Rahman, editor Studies in natural products chemistry: bioactive natural products. Vol. 33. Amsterdam; Elsevier Science Publishers 2006: 141-92
• 26 Fernandes J, Castilho R O, Costa M R, Wagner-Souza K, Kaplan M AC, Gattass C R. Pentacyclic triterpenes from Chrysobalanaceae species: cytotoxicity on multidrug resistant and sensitive leukemia cell lines.  Cancer Lett. 2003;  190 165-9
• 27 Yoshida M, Fuchigami M, Nagao T, Okave H, Matsunaga K, Takata J. et al . Antiproliferative constituents from Umbelliferae plants VII. Active triterpenes and rosmaric acid from Centella asiatica.  Biol Pharm Bull. 2005;  28 173-5
• 28 Neto C, Vaisberg A, Zhou B, Kingston D, Hammond G. Cytotoxic triterpene acids the peruvian plant Polylepis racemosa.  Planta Med. 2000;  66 483-4
• 29 Vasconcelos F C, Gattass C R, Rumjanek V M, Maia R C. Pomolic acid-induced apoptosis in cells from patients with chronic myeloid leukemia exhibiting different drug resistance profile.  Invest New Drugs. 2007;  25 5253-33
• 30 Fernandes J, Weinlich R, Castilho R O, Kaplan M A, Amarante-Mendes G P, Gattass C R. Pomolic acid triggers mitochondria-dependent apoptotic cell death in leukemia cell line.  Cancer Lett. 2005;  219 49-55
• 31 Fernandes J, Weinlich R, Castilho R O, Amarante-Mendes G P, Gattass C R. Pomolic acid may overcome multidrug resistance mediated by overexpression of anti-apoptotic Bcl-2 proteins.  Cancer Lett. 2007;  245 315-20