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
Key words
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.
13 C-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
José-Luis Rios
Departament de Farmacologia
Facultat de Farmàcia
Universitat de València
Av. Vicent Andrés Estellés s/n
46100 Burjassot (Valencia)
Spain
Phone: +34-963-544-973
Fax: +34-963-544-498
Email: riosjl@uv.es