Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
BMC Complementary Medicine and Therapies
Published in: BMC Complementary Medicine and Therapies 1/2015

Open Access 01-12-2015 | Research article

The effect of 3β, 6β, 16β-trihydroxylup-20(29)-ene lupane compound isolated from Combretum leprosum Mart. on peripheral blood mononuclear cells

Authors: Fabianne Lacouth-Silva, Caroline V. Xavier, Sulamita da S. Setúbal, Adriana S. Pontes, Neriane M. Nery, Onassis Boeri de Castro, Carla F. C. Fernandes, Eduardo R. Honda, Fernando B. Zanchi, Leonardo A. Calderon, Rodrigo G. Stábeli, Andreimar M. Soares, Izaltina Silva-Jardim, Valdir A. Facundo, Juliana P. Zuliani

Published in: BMC Complementary Medicine and Therapies | Issue 1/2015

Login to get access

Abstract

Background

The Combretum leprosum Mart. plant, popularly known as mofumbo, is used in folk medicine for inflammation, pain and treatment of wounds. From this species, it is possible to isolate three triterpenes: (3β, 6β, 16β-trihydroxylup-20(29)-ene) called lupane, arjunolic acid and molic acid. In this study, through preclinical tests, the effect of lupane was evaluated on the cytotoxicity and on the ability to activate cellular function by the production of TNF-α, an inflammatory cytokine, and IL-10, an immuno regulatory cytokine was assessed. The effect of lupane on the enzymes topoisomerase I and II was also evaluated.

Methods

For this reason, peripheral blood mononuclear cells (PBMCs) were obtained and cytotoxicity was assessed by the MTT method at three different times (1, 15 and 24 h), and different concentrations of lupane (0.3, 0.7, 1.5, 6, 3 and 12 μg/mL). The cell function was assessed by the production of TNF-α and IL-10 by PBMCs quantified by specific enzyme immunoassay (ELISA). The activity of topoisomerases was assayed by in vitro biological assays and in silico molecular docking.

Results

The results obtained showed that lupane at concentrations below 1.5 μg/mL was not toxic to the cells. Moreover, lupane was not able to activate cellular functions and did not alter the production of IL-10 and TNF-α. Furthermore, the data showed that lupane has neither interfered in the action of topoisomerase I nor in the action of topoisomerase II.

Conclusion

Based on preclinical results obtained in this study, we highlight that the compound studied (lupane) has moderate cytotoxicity, does not induce the production of TNF-α and IL-10, and does not act on human topoisomerases. Based on the results of this study and taking into consideration the reports about the anti-inflammatory and leishmanicidal activity of 3β, 6β, 16β-trihydroxylup-20(29)-ene, we suggest that this compound may serve as a biotechnological tool for the treatment of leishmaniasis in the future.
Literature
1.
Agra MF, Freitas PF, Barbosa-Filho JM. Synopsis of the plants known as medicinal and poisonous in Northeast of Brazil. Rev Bras Farmacogn. 2007;17:114–40.CrossRef
2.
Hoareau L, Da Silva EJ. Medicinal plants: A re-emerging health aid. Eletronic J Biotechnol. 1999;2:56–70.
3.
Sneader W. Drug Prototypes and Their Exploitation. United Kingdom: Wiley; 1996.
4.
Calderon LA, Silva-Jardim I, Zuliani JP, Silva AA, Ciancaglini P, Pereira-da-Silva LH, et al. Amazonian biodiversity: a view of drug development for Leishmaniasis and Malaria. J Braz Chem Soc. 2009;20:1011–23.CrossRef
5.
Tan F, Shi S, Zhong Y, Gong X, Wang Y. Phylogenetic relationships of Combretoideae (Combretaceae) inferred from plastid, nuclear gene and spacer sequences. Journal of Plant Research. 2002;115:475–81.CrossRefPubMed
6.
Pettit GR, Singh SB, Boyd MR, Hamel E, Pettit RK, Schmidt JM, et al. Antineoplastic agents. 291. Isolation and synthesis of combretastatins A-4, A-5, and A-6(1a). Med Chem. 1995;38:1666–72.CrossRef
7.
Facundo VA, Andrade CHS, Silveira ER, Braz-Filho R, Hufford C. Triterpenes and flavonoids from Combretum leprosum. Phytochesmistry. 1993;1993(32):411–5.CrossRef
8.
Pio Correa M. Diccionário de Plantas Úteis do Brasil e das Exóticas Cultivadas, vol. 2. Rio de Janeiro: Imprensa Nacional; 1984.
9.
Lira SRD, Almeida RN, Almeida FRC, Oliveira FS, Duarte JC. Preliminary studies on the analgesic properties of the ethanol extract of C. leprosum. Pharm Biol. 2002;40:213–5.CrossRef
10.
Pietrovsky EF, Rosa KA, Facundo VA, Rios K, Marques MCA, Santos ARS. Antinociceptive properties of the ethanolic extract and of the triterpene 3β, 6β, 16β-trihidroxilup-20(29)-ene obtained from the flowers of Combretum leprosum in mice. Pharmacology, Biochemistry and Behavior. 2006;83:90–9.CrossRef
11.
Fernandes FF, Tomaz MA, El-Kik CZ, Monteiro-Machado M, Strauch MA, Cons BL, et al. Counteraction of Bothrops snake venoms by Combretum leprosum root extract and arjunolic acid. J Ethnopharmacol. 2014;155:552–62.CrossRefPubMed
12.
Evaristo FF, Albuquerque MR, Dos Santos HS, Bandeira PN, Avila Fdo N, Da Silva R, et al. Antimicrobial effect of the triterpene 3β,6β,16β-trihydroxylup-20(29)-ene on planktonic cells and biofilms from Gram positive and Gram negative bacteria. Biomed Res Int. 2014;2014:729358. doi:10.​1155/​2014/​729358.PubMedCentralCrossRefPubMed
13.
Teles CBG, Moreira LS, Silva AAE, Facundo VA, Zuliani JP, Stábeli RG, et al. Activity of the Lupane isolated from Combretum leprosum against Leishmania amazonensis promastigotes. J Braz Chem Soc. 2011;22:936–42.CrossRef
14.
Nagase M, Oto J, Sugiyama S, Yube K, Takaishi Y, Sakato N. Apoptosis induction in HL-60 cells an inhibition of topoisomerase II by triterpene celastrol. Biosci Biotechnol Biochem. 2003;67:1883–7.CrossRefPubMed
15.
Sethi G, Ahn KS, Pandey MK, Aggarwal BB. Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-kappaB-regulated gene products and TAK-1-mediated NF-kappaB activation. Blood. 2007;109:2727–35.PubMed
16.
Wang P, Ownby S, Zhang Z, Yuan W, Li S. Cytotoxicity and inhibiton of DNA topoisomerase I of polyhydroxyleted triterpenoids and triterpenoid glycosides. Chem lett. 2010;20:2790–6.CrossRef
17.
Chowdhury AR, Mandal S, Goswami A, Ghosh M, Mandal L, Chakraborty D, et al. Dihydrobetulinic acid induces apoptosis in Leishmania donovani by targeting DNA topoisomerase I and II: implications in antileishmanial therapy. Mol Med. 2003;9:26–36.PubMedCentralPubMed
18.
Chowdhury S, Mukherjee T, Sengupta S, Chowdhury SR, Mukhopadhyay S, Majumder HK. Novel betulin derivatives as antileishmanial agents with mode of action targeting type IB DNA topoisomerase. Mol Pharmacol. 2011;80:694–703.CrossRefPubMed
19.
Reilly TP, Bellevue FH, Woster PM. Compararion of the in vitro citotoxicity os hydroxylamine metabolites of sulfamethoxazole and dapsone. Biochem Pharmacol. 1998;55:803–10.CrossRefPubMed
20.
Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK, Goodsell DS, et al. AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility. J Comp Chem. 2009;30:2785–91.CrossRef
21.
Ioanoviciu A, Antony S, Pommier Y, Staker BL, Stewart L, Cushman M. Synthesis and mechanism of action studies of a series of norindenoisoquinoline topoisomerase I poisons reveal an inhibitor with a flipped orientation in the ternary DNA-enzyme-inhibitor complex as determined by X-ray crystallographic analysis. J Med Chem. 2005;48:4803–14.CrossRefPubMed
22.
Wu CC, Li TK, Farh L, Lin LY, Lin TS, Yu YJ, et al. Structural basis of type II topoisomerase inhibition by the anticancer drug etoposide. Science. 2011;333:459–62.CrossRefPubMed
23.
Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, Ferrin TE, et al. UCSF Chimera - a visualization system for exploratory research and analysis. J Comput Chem. 2004;25:1605–12.CrossRefPubMed
24.
Morais-Lima GR, De Sales IR, Caldas-Filho MR, De Jesus NZ, De Sousa FH, Barbosa-Filho JM, et al. Bioactivities of the genus Combretum (Combretaceae): a review. Molecules. 2012;17:9142–206.CrossRefPubMed
25.
Jesus JA, Lago JH, Laurenti MD, Yamamoto ES, Passero LF. Antimicrobial activity of oleanolic and ursolic acids: an update. Evid Based Complement Alternat Med. 2015;2015:1–14.
26.
Thimmappa R, Geisler K, Louveau T, O’Maille P, Osbourn A. Triterpene biosynthesis in plants. Annu Rev Plant Biol. 2014;65:225–57.CrossRefPubMed
27.
Kommera H, Kaluđerović GN, Kalbitz J, Paschke R. Lupane triterpenoids--betulin and betulinic acid derivatives induce apoptosis in tumor cells. Invest New Drugs. 2011;29:266–72.CrossRefPubMed
28.
Laszczyk MN. Pentacyclic triterpenes of the lupane, oleanane and ursane group as tools in cancer therapy. Planta Med. 2009;75:1549–60.CrossRefPubMed
29.
Baltina LA, Flekhter OB, Nigmatullina LR, Boreko EI, Pavlova NI, Nikolaeva SN, et al. Lupane triterpenes and derivatives with antiviral activity. Bioorg Med Chem Lett. 2003;13:3549–52.CrossRefPubMed
30.
De Silva ML, David JP, Silva LC, Santos RA, David JM, Lima LS, et al. Bioactive oleanane, lupane and ursane triterpene acid derivatives. Molecules. 2012;17:12197–205.CrossRefPubMed
31.
Fulda S. Betulinic acid: A natural product with anticancer activity. Mol Nutr Food Res. 2009;53:140–6.CrossRefPubMed
32.
Fulda S, Friesen C, Los M, Scaffidi C, Mier W, Benedict M, et al. Betulinic acid triggers CD95 (APO-1/Fas)- and p53-independent apoptosis via activation of caspases in neuroectodermal tumors. Cancer Res. 1997;57:4956–64.PubMed
33.
Fulda S, Debatin KM. Betulinic acid induces apoptosis through a direct effect on mitochondria in neuroectodermal tumors. Med Pediatr Oncol. 2000;35:616–8.CrossRefPubMed
34.
Galaĭko NV, Tolmacheva IA, Grishko VV, Volkova LV, Prevozchikova EN, Pestereva SA. Antiviral activity of 2, 3-secotriterpenic hydrazones of lupane and 19 beta, 28-epoxy-18 alpha-oleanane type. Bioorganicheskaya Khimiya. 2010;36:556–62.PubMed
35.
Kwon HJ, Shim JS, Kim JH, Cho HY, Yum YN, Kim SH, et al. Betulinic acid inhibits growth factor-induced in vitro angiogenesis via the modulation of mitochondrial function in endothelial cells. Jpn J Cancer Res. 2002;93:417–25.CrossRefPubMed
36.
Liaw CC, Chen YC, Huang GJ, Tsai YC, Chien SC, Wu JH, et al. Anti-inflammatory lanostanoids and lactone derivatives from Antrodia camphorata. J Nat Prod. 2013;76:489–94.CrossRefPubMed
37.
Melzig MF, Bormann H. Betulinic acid inhibits aminopeptidase N activity. Planta Med. 1998;64:655–7.CrossRefPubMed
38.
Mokoka TA, McGaw LJ, Mdee LK, Bagla VP, Iwalewa EO, Eloff JN. Antimicrobial activity and cytotoxicity of triterpenes isolated from leaves of Maytenus undata (Celastraceae). BMC Complement. Altern. Med. 2013, 13, doi:10.​1186/​1472-6882-13-111
39.
Basu A, Krishnamurthy S. Cellular responses to Cisplatin-induced DNA damage. J Nucleic Acids. 2010;2010:1–16.
40.
Sedletska Y, Giraud-Panis MJ, Malinge JM. Cisplatin is a DNA-damaging antitumour compound triggering multifactorial biochemical responses in cancer cells: importance of apoptotic pathways. Current Medicinal Chemistry Anti-Cancer Agents. 2005;5:251–65.CrossRefPubMed
41.
Florea AM, Büsselberg D. Cisplatin as an anti-tumor drug: cellular mechanisms of activity, drug resistance and induced side effects. Cancers (Basel). 2011;3:1351–71.CrossRef
42.
Rademaker-Lakhai JM, Crul M, Zuur L, Baas P, Beijnen JH, Simis YJ, et al. Relationship between cisplatin administration and the development of ototoxicity. J Clin Oncol. 2006;24:918–24.CrossRefPubMed
43.
Tsang RY, Al-Fayea T, Au HJ. Cisplatin overdose: Toxicities and management. Drug Saf. 2009;32:1109–22.CrossRefPubMed
44.
Hartmann JT, Kollmannsberger C, Kanz L, Bokemeyer C. Platinum organ toxicity and possible prevention in patients with testicular cancer. Int J Cancer. 1999;83:866–9.CrossRefPubMed
45.
Shen FZ, Wang J, Liang J, Mu K, Hou JY, Wang YT. Low-dose metronomic chemotherapy with cisplatin: can it suppress angiogenesis in H22 hepatocarcinoma cells? Int J Exp Pathol. 2010;91:10–6.PubMedCentralCrossRefPubMed
46.
Emmerich D, Vanchanagiri K, Baratto LC, Schmidt H, Paschke R. Synthesis and studies of anticancer properties of lupane-type triterpenoid derivatives containing a cisplatin fragment. Eur J Med Chem. 2014;75:460–6.CrossRefPubMed
47.
Teles CB, Moreira-Dill LS, Silva A de A, Facundo VA, de Azevedo Jr WF, da Silva LH, et al. A lupane-triterpene isolated from Combretum leprosum Mart. fruit extracts that interferes with the intracellular development of Leishmania (L.) amazonensis in vitro. BMC Complement Altern Med. 2015;15:165. doi:10.​1186/​s12906-015-0681-9.PubMedCentralCrossRefPubMed
48.
Koster DA, Croquette V, Dekker C, Shuman S, Dekker NH. Friction band torque govern the relaxation of DNA supercoils by eukaryotic topoisomerase IB. Nature. 2005;434:671–4.CrossRefPubMed
49.
Liu LF, Liu CC, Alberts BM. Type II DNA topoisomerases: enzymes that can unknot a topologically knotted DNA molecule via a reversible double-strand break. Cell. 1980;19:697–707.CrossRefPubMed
50.
Champoux JJ. DNA topoisomerases: structure, function, and mechanism. Annu Rev Biochem. 2001;70:369–413.CrossRefPubMed
51.
Wang JC. Cellular roles of DNA topoisomerases: a molecular perspective. Nat Rev Mol Cell Biol. 2002;3:430–40.CrossRefPubMed
52.
D’Arpa P, Machlin PS, Ratrie H, Rothfield NF, Cleveland DW, Earnshaw WC. cDNA cloning of human DNA topoisomerase I: catalytic activity of a 67.7-kDa carboxyl-terminal fragment. Proc Natl Acad Sci USA. 1988;85:2543–7.PubMedCentralCrossRefPubMed
53.
Baker SD, Wadkins RM, Stewart CF, Beck WT, Danks MK. Cell cycle analysis of amount and distribution of nuclear DNA topoisomerase I as determined by fluorescence digital imaging microscopy. Cytometry. 1995;19:134–45.CrossRefPubMed
54.
Meyer KN, Kjeldsen E, Straub T, Knudsen BR, Hickson ID, Kikuchi A, et al. Cell cycle-coupled relocation of types I and II topoisomerases and modulation of catalytic enzyme activities. J Cell Biol. 1997;136:775–88.PubMedCentralCrossRefPubMed
55.
Christensen MO, Barthelmes HU, Feineis S, Knudsen BR, Andersen AH, Boege F, et al. Changes in mobility account for camptothecin-induced subnuclear relocation of topoisomerase I. J Biol Chem. 2002;277:15661–5.CrossRefPubMed
56.
Beldner MA, Sherman CA, Green MR, Garrett-Mayer E, Chaudhary U, Meyer ML, et al. Phase I dose escalation study of vinorelbine and topotecan combination chemotherapy in patients with recurrent lung cancer. BMC Cancer. 2007;7:231–45.PubMedCentralCrossRefPubMed
57.
Lim WT, Baggstrom MQ, Read W, Fracasso PM, Govindan R. A Phase I trial of weekly docetaxel and topotecan for solid tumors. Acta Oncol. 2008;47:311–5.CrossRefPubMed
58.
Friedman HS, Keir ST, Houghton PJ. The emerging role of irinotecan (CPT-11) in the treatment of malignant glioma in brain tumors. Cancer. 2003;97:2359–62.CrossRefPubMed
59.
Han JY, Lim HS, Park YH, Lee SY, Lee JS. Integrated pharmacogenetic prediction of irinotecan pharmacokinetics and toxicity in patients with advanced non-small cell lung cancer. Lung Cancer. 2009;63:115–20.CrossRefPubMed
60.
Hsiang YH, Hertzberg R, Hecht S, Liu LF. Camptothecin induces protein-linked DNA breaks via mammalian DNA topoisomerase I. J Biol Chem. 1985;260:14873–8.PubMed
61.
Drake FH, Zimmerman JP, McCabe FL, Bartus HF, Per SR, Sullivan DM, et al. Purification of topoisomerase II from Amsacrine-resistant P388 leukemia cells. Evidence for two forms of the enzyme. J Biol Chem. 1987;262:16739–47.PubMed
62.
Krieser RJ, Maclea KS, Park JP, Eastman A. The cloning, genomic structure, localization, and expression of human deoxyribonuclease IIbeta. Gene. 2001;2001(269):205–16.CrossRef
63.
Linka RM, Porter AC, Volkov A, Mielke C, Boege F, Christensen MO. C-terminal regions of topoisomerase IIalpha and IIbeta determine isoform-specific functioning of the enzymes in vivo. Nucleic Acids Res. 2007;35:3810–22.PubMedCentralCrossRefPubMed
64.
Asano T, Kleinerman ES, Zwelling LA, Zhou Z, Fukunaga Y. Adenovirus-mediated human topoisomerase IIalpha gene transfer increases the sensitivity of etoposide-resistant human and mouse breast cancer cells. Acta Oncol. 2005;44:240–7.CrossRefPubMed
65.
66.
Kellner U, Rudolph P, Parwaresch R. Human DNA-topoisomerase – diagnostic and therapeutic implications for cancer. Onkologie. 2000;23:424–30.CrossRefPubMed
67.
68.
Bodley AL, Chakraborty AK, Xie S, Burri C, Shapiro TA. An unusual type IB topoisomerase from African trypanosomes. Proc Natl Acad Sci USA. 2003;100:7539–44.PubMedCentralCrossRefPubMed
69.
Das BB, Sen N, Ganguly A, Majumder HK. Reconstitution and functional characterization of the unusual bi-subunit type I DNA topoisomerase from Leishmania donovani. FEBS Lett. 2004;565:81–8.CrossRefPubMed
70.
Reguera RM, Redondo CM, Gutierrez De Prado R, Pérez-Pertejo Y, Balaña-Fouce R. DNA topoisomerase I from parasitic protozoa: a potential target for chemotherapy. Biochim Biophys Acta. 2006;1759:117–31.CrossRefPubMed
71.
Villa H, Otero Marcos AR, Reguera RM, Balaña-Fouce R, García-Estrada C, Pérez-Pertejo Y, et al. A novel active DNA topoisomerase I in Leishmania donovani. J Biol Chem. 2003;278:3521–6.CrossRefPubMed
Metadata
Title
The effect of 3β, 6β, 16β-trihydroxylup-20(29)-ene lupane compound isolated from Combretum leprosum Mart. on peripheral blood mononuclear cells
Authors
Fabianne Lacouth-Silva
Caroline V. Xavier
Sulamita da S. Setúbal
Adriana S. Pontes
Neriane M. Nery
Onassis Boeri de Castro
Carla F. C. Fernandes
Eduardo R. Honda
Fernando B. Zanchi
Leonardo A. Calderon
Rodrigo G. Stábeli
Andreimar M. Soares
Izaltina Silva-Jardim
Valdir A. Facundo
Juliana P. Zuliani
Publication date
01-12-2015
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2015
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-015-0948-1

Other articles of this Issue 1/2015

BMC Complementary Medicine and Therapies 1/2015 Go to the issue

Research article

Hypoglycemic and hypolipidemic activities of aqueous extract of flowers from Nycantus arbor-tristis L. in male mice

Research article

Antioxidant potential, total phenolic and total flavonoid contents of Rhododendron anthopogonoides and its protective effect on hypoxia-induced injury in PC12 cells

Research article

The anti-inflammatory activities of ethanol extract from Dan-Lou prescription in vivo and in vitro

Research article

Comparison of the efficacy of aroma-acupressure and aromatherapy for the treatment of dementia-associated agitation

Study protocol

Immediate changes in electroencephalography activity in individuals with nonspecific chronic low back pain after cranial osteopathic manipulative treatment: study protocol of a randomized, controlled crossover trial

Research article

Baicalein reduces the occurrence of cirrhotic endotoxemia by reducing intestinal mucosal apoptosis