CHAPTER 7: THE HOUSEHOLD DIVISION OF LABOUR
7.2. WOMEN HAVE RESPONSIBILITY FOR HOUSEHOLD LABOUR
Para a realização das análises histopatológicas, as patas infectadas dos camundongos foram coletadas após 2 e 8 semanas de tratamento com extrato hidroalcoólico de própolis verde, correspondentes à 4 e 10 semanas de infecção, respectivamente.
Os resultados das análises revelaram que, após 4 semanas de infecção (2 semanas de tratamento), os animais tratados com o extrato hidroalcoólico da própolis verde exibiram intenso infiltrado inflamatório, de distribuição difusa, composto de forma majoritária por macrófagos. Ao passo que no grupo controle a inflamação foi discreta a ausente. Em ambos os grupos a epiderme apresentava-se íntegra em todas as camadas, bem como a derme, com seus respectivos anexos cutâneos de aspecto e distribuição convencional.
Na décima semana de infecção (8 semanas de tratamento), observou-se um intenso infiltrado inflamatório na derme, em ambos os grupos, controle e tratado, caracterizado por um perfil inflamatório crônico, distribuído em toda a camada reticular da derme, de forma difusa, caracterizado de forma predominante pela presença de macrófagos, linfócitos e esparsos eosinófilos. Foram observados ainda, alguns macrófagos com citoplasma amplo, de aspecto vacuolizado (Figura 13). Desta forma, as análises histopatológicas demonstraram que o extrato hidroalcoólico da própolis foi capaz de recrutar de forma precoce os macrófagos para o sítio da lesão causada por L. amazonensis.
RESULTADOS E DISCUSSÃO
51
Figura 13: Fotomicrografia de cortes histológicos (4 μm, HE) de coxim plantar de camundongos C57BL/6 infectados por L. amazonensis. Figuras A e C indicam os grupos controle infectados e não tratados, por 2 e 8 semanas, respectivamente. Figuras B e D indicam os grupos infectados e tratados por 2 e 8 semanas, respectivamente. Aumento 40x. As setas indicam infiltrado inflamatório difuso na derme.
Estes resultados são condizentes com dados da literatura, que apontam que a própolis modula a imunidade não específica por meio do recrutamento de macrófagos. Tatefuji et al. (1996) demonstraram que seis compostos isolados da própolis, identificados como ácidos cafeoilquínicos, são capazes de aumentar a motilidade e distribuição dos macrófagos. Da mesma forma, Miranda et al. (2015) observaram que camundongos BALB/c infectados por L.
amazonensis e tratados com própolis na dose de 5 mg/kg/dia apresentaram um significativo aumento no infiltrado de macrófagos no sítio da lesão quando comparados com o grupo controle infectado.
O fato de o extrato hidroalcoólico da própolis verde ter aumentado o recrutamento de macrófagos para o sítio da infecção aliado à possível inibição da produção de IFN-γ por estas células tornou o ambiente favorável ao desenvolvimento do parasito, deixando o organismo susceptível à infecção.
RESUMO DOS RESULTADOS
52
7RESUMO DOS RESULTADOS
Neste estudo, os efeitos leishmanicidas do extrato hidroalcoólico da própolis verde foram investigados in vitro, em formas promastigotas, amastigotas axênicas e amastigotas intracelulares de L. amazonensis e in vivo, em camundongos C57BL/6 infectados por L.
amazonensis.
Observou-se que este extrato não possui efeitos citotóxicos sobre macrófagos peritoneais murinos e apresenta atividade inibitória sobre todas as formas de L. amazonensis testadas in vitro, de maneira concentração-dependente. Enquanto que in vivo, para a dose testada, foram observados efeitos leishmanicidas apenas nos estágios iniciais da infecção, havendo até mesmo aumento da carga parasitária no linfonodo poplíteo drenante no estágio mais tardio. Além disso, o tratamento não teve influência significativa no tamanho da lesão apresentada pelos animais, havendo apenas uma tendência a ser menor quando comparada com o grupo controle.
A partir destes resultados, conclui-se que a própolis pode representar uma potencial alternativa terapêutica para utilização em associação no tratamento da leishmaniose. No entanto, são necessárias mais investigações a respeito da sua atividade leishmanicida in vivo, principalmente. Destaca-se a necessidade de testar outros regimes terapêuticos, com diferentes doses e vias de administração. Além disso, é preciso avaliar quais são os efeitos leishmanicida e imunomodulador dos diferentes extratos da própolis (alcoólico, aquoso e hidroalcoólico), a fim de elucidar melhor tais efeitos de cada tipo de extrato e esclarecer até que ponto a imunomodulação exercida pela própolis influencia na atividade leishmanicida dos diferentes tipos de extrato de própolis.
CONCLUSÃO
53
8CONCLUSÃO
Conclui-se que o extrato hidroalcoólico da própolis verde (Cytopropolis®) possui efeitos inibitórios sobre L. amazonensis in vitro, de maneira concentração-dependente. E, para a infecção in vivo, este extrato possui efeito leishmanicida apenas nos estágios iniciais da infecção. Além disso, por se tratar de um extrato que é submetido à um rigoroso controle de qualidade, há uma garantia da confiabilidade e reprodutibilidade dos resultados obtidos neste trabalho.
REFERÊNCIAS
54
REFERÊNCIAS
AFONSO, L.C. & SCOTT, P. Immune responses associated with susceptibility of C57BL/10 mice to Leishmania amazonensis. Infect. Immun. v. 61, p. 2952-2959, 1993.
AKAO, Y. et al. Cell growth inhibitory effect of cinnamic acid derivatives from propolis on human tumor cell lines. Biol Pharm Bull. v.26(7), p.1057-9, 2003.
ALDAY, E.; VALENCIA, D.; CARREÑO, AN.; PICERNO, P.; PICCINELLI, AL.; RASTRELLI, L.; ROBLES-ZEPEDA, R.; HERNANDEZ, J.; VELAZQUEZ, C. Apoptotic induction by pinobanksin and some of its ester derivatives from Sonoran propolis in a B-cell lymphoma cell line. Chemico-Biological Interactions. v. 242, p. 35–445, 2015.
ALEXANDER, J. & BROMBACHER, F. T helper1/T helper2 cells and resistance/susceptibility to Leishmania infection: is this paradigm still relevant? Frontiers in
immunology. v. 3, 2012.
ALEXANDER, J. & BRYSON, K. T helper (h)1/Th2 and Leishmania: paradox rather than paradigm. Immunology Letters. v. 99, p. 17–23, 2005.
ANSORGE, S.; REINHOLD, D.; LENDECKEL, U. Propolis and some of its constituents down-regulate DNA synthesis and inflammatory cytokine production but induce TGF-beta 1 production of human immune cells. Zeitschrift fur Naturforschung. v. 58c, p. 580–589, 2003.
ANTOINE, J.C.; PRINA, E.; LANG, T.; COURRET, N. The biogenesis and properties of the parasitophorous vacuoles that harbour Leishmania in murine macrophages. Trends
microbiol. v. 07, n. 20, 1998.
ARVOUET-GRAND, A.; LEJEUNE, B.; BASTIDE, P.; POURRAT, A.; PRIVAT, AM.; LEGRET, P. Propolis extract. I. Acute toxicity and determination of acute primary cutaneous irritation index. J Pharm Belg. v. 48 (3), p. 165-70, 1993.
AWASTHI, A.; MATHUR, R. K.; SAHA, B. Immune response to Leishmania infection.
Indian J. Med. Res. v. 119 (6), p. 238-258, 2004.
AYRES, D.C.; MARCUCCI, M.C.; GIORGIO, S. Effects of Brazilian propolis on
Leishmania amazonensis. Mem Inst Oswaldo Cruz. v. 102, n. 2, p. 215 – 220, 2007.
BAHIA, MT.; NASCIMENTO, AF.; MAZZETI, AL.; MARQUES, LF.; GONÇALVES, KR.; MOTA, LW.; DINIZ, LF.; CALDAS, IS.; TALVANI, A.; SHACKLEFORD, DM.; KOLTUN, M.; SAUNDERS, J.; WHITE, KL.; SCANDALE, I.; CHARMAN, SA.; CHATELAIN, E. Antitrypanosomal activity of fexinidazole metabolites, potential new drug candidates for Chagas disease. Antimicrob Agents Chemother. v. 58(8), p. 4362-70, 2014. BANKOVA, V. Chemical diversity of propolis and the problem of standardization. Journal
of Ethnopharmacology. v. 100, p. 114–117, 2005a.
BANKOVA,V. Recent trends and important developments in propolis research. Evid Based
REFERÊNCIAS
55 BURDOCK, GA. Review of the biological properties and toxicity of bee propolis (propolis).
Food Chem Toxicol. v. 36, p. 347–363, 1998.
CARVALHO, P.B. & FERREIRA, E.I. Leishmaniais phytotherapy. Nature’s leadership against na ancient disease. Fitoterapia. v. 72, p.599-618, 2001.
CHAKRABORTY, A.K.; MAJUMDER, H.K. Mode of action of pentavalent antimonials: specific inhibition of type I DNA topoisomerase of Leishmania donovani. Biochem.
Biophys. Res. Commun. v. 152, p. 605-611, 1988.
CHAN, G.C.F.; CHEUNG, K.W.; SZE, D.M.Y. The Immunomodulatory and Anticancer Properties of Propolis. Clinic Rev Allerg Immunol. v. 44, p.262–273, 2013.
CROSS, A. S.; SAKARYA, S. et al. Recruitment of murine neutrophils in vivo through endogenous sialidase activity. J. Biol. Chem, v.278, n.6, Feb 7, p.4112-20, 2003.
CUNNINGHAM, A. C. Parasitic adaptive mechanisms in infection by Leishmania. Exp Mol
Pathol. v. 72, p. 132-141, 2002.
DANTAS, AP.; SALOMÃO, K.; BARBOSA, HS.; DE CASTRO, SL. The effect of Bulgarian propolis against Trypanosoma cruzi and during its interaction with host cells. Men
Inst Oswaldo Cruz. v. 101, p. 207-211, 2006.
DENTON, H.; MCGREGOR, J.C.; COOMBS, G.H. Reduction of anti-leishmanial pentavalent antimonial drugs by a parasite-specific thiol-dependent reductase, TDR1.
Biochem. J. v. 381, p. 405-412, 2004.
FERREIRA, C.S.; MARTINS, P.S.; DEMICHELI, C.; BROCHU, C.; OUELLETTE,M.; FRÉZARD, F. Thiol-induced reduction of antimony(V) into antimony(III): a comparative study with trypanothione, cysteinylglycine, cysteine and glutathione. BioMetals. v. 16, p. 441-443, 2003.
FERREIRA, F.M.; CASTRO, R.A.O.; BATISTA, M.A.; ROSSI, F.M.D.; SILVEIRA- LEMOS, D.; FRÉZARD, F.; MOURA, S.A.L.; REZENDE, S.A. Association of water extract of green propolis and liposomal meglumine antimoniate in the treatment of experimental visceral leishmaniasis. Parasitol Res. v. 113, p. 533–543, 2014.
FREITAS, SF.; SHINOHARA, L.; SFORCIN, JM., GUIMARÃES, S. In vitro effects of propolis on Giardia duodenalis trophozoites. Phytomedicine. v. 13, p. 170-175, 2006.
FRÉZARD, F.; DEMICHELI, C.; FERREIRA, C.S.; COSTA, M.A.P. Glutathione-induced conversion of pentavalent antimony to trivalent antimony in meglumine antimoniate.
Antimicrob. Agents Chemother. v. 45, p. 913-916, 2001.
FRÉZARD, F.; DEMICHELI, C.; RIBEIRO, R.R. Pentavalent antimonials: new perspectives for old drugs. Molecules. v. 14 (7), p. 2317-36, 2009.
GOURBAL, B.; SONUC, N.; BHATTACHARJEE, H.; LEGARE, D.; SUNDAR, S.; OUELLETTE, M.; ROSEN, B.P.; MUKHOPADHYAY, R. Drug uptake and modulation of drug resistance in Leishmania by an aquaglyceroporin. J. Biol. Chem. v. 279, p. 31010- 31017, 2004.
REFERÊNCIAS
56 GRIMALDI, J.R.G.; DAVID, J.R.; MCMAHON-PRATT, D. Identification and distribution of New World Leishmania species caracterized by serademe analysis using monoclonal antibodies. The American Journal of Tropical Medicine and Hygiene, v. 36, p. 270-287, 1987.
GUPTA, N.; GOYAL, N.; RASTOGI, AK. In vitro cultivation and characterization of axenic amastigotes of Leishmania. Trends in Parasitology. v.17, n. 3, 2001.
HANDLER, M.Z.; PATEL, P.A.; KAPILA, R.; AL-QUBATI, Y.; SCHWARTZ, R.A. Cutaneous and mucocutaneous leishmaniasis: Clinical perspectives. J Am Acad Dermatol. v. 73(6), p. 897-908, 2015.
HARMS, G.; SCHONIAN, G.; FELDMEIER, H. Leishmaniasis in Germany. Emerg infect
dis. v. 9, n. 7, p. 872-5, 2003.
HIGASHI, KO.; DE CASTRO, SL. Propolis extracts are effective against Trypanosoma cruzi and have an impact on its interaction with host cells. J Ethnopharmacol. v. 43, p. 149-155, 1994.
HU, F.; HEPBURN, H.R.; LI, Y.; CHEN, M.; RADLOFF, S.E.; DAYA, S. Effects of ethanol and water extracts of propolis (bee glue) on acute inflammatory animal models. Journal of
Ethnopharmacology. v. 100, p. 276–283, 2005.
INTERNATIONAL ORGANIZATION FOR STANDARDIZATION (ISO). Biological evaluation of medical devices — Part 5: Tests for in vitro cytotoxicity. ISO 2009, p. 9 -28, ISO 2009.
JOSHI, P. B.; KELLY, B. L.; KAMHAWI, S.; SACKS, D. L. & MCMASTER, W. R. Targeted gene deletion in Leishmania major identifies leishmanolysin (GP63) as a virulence factor. Mol. Biochem. Parasitol. v. 120, p. 33–40, 2002.
KANSAL, S.; TANDON, R.; DWIVEDI, P.; MISRA, P.; VERMA, P. R. P.; DUBE, A.; MISHRA, P.R. Development of nanocapsules bearing doxorubicin for macrophage targeting through the phosphatidylserine ligand: a system for intervention in visceral leishmaniasis. J
Antimicrob Chemother. v. 67, p. 2650–2660, 2012.
KAYE, P. e SCOTT, P. Leishmaniasis: complexity at the host–pathogen interface. Nat Rev
Microbiol.v.9(8), p. 604-15, 2011.
KEVRIC, I.; CAPPEL, M.A.; KEELING, J.H. New World and Old World Leishmania Infections: A Practical Review. Dermatol Clin. v.33, n. 3, p. 579-93, 2015.
LAINSON, R. The American leishmaniases: some observations on their ecology and epidemiology. Trans.R.Soc.Trop.Med.Hyg., v. 77, n. 5, p. 569-596, 1983.
LIMA, LD.; ANDRADE, SP.; CAMPOS, PP.; BARCELOS, LS.; SORIANI, FM.; MOURA. SA.; FERREIRA, MA. Brazilian green propolis modulates inflammation, angiogenesis and fibrogenesis in intraperitoneal implant in mice. BMC Complement Altern Med. v. 14, p. 177, 2014.
REFERÊNCIAS
57 LOEUILLET, C.; BAÑULS, AL.; HIDE, M. Study of Leishmania pathogenesis in mice: experimental considerations. Parasites & Vectors. v. 9, p.144, 2016.
LUPI, O.; BARTLETT, B.L.; HAUGEN, R.N. et al. Tropical dermatology: Tropical diseases caused by protozoa. J am Acad dermatol. v. 60, n. 6, p. 897-925, 2009.
MALTEZOU HC. Visceral leishmaniasis: advances in Treatment. Recent Pat Antiinfect
Drug Discov. v. 3, n. 3, p.192-8, 2008.
MARCUCCI, M. C. Propolis: chemical composition, biological properties and therapeutic activity. Apidologie. v. 26, n. 2, p. 83-99, 1995.
MARCUCCI, M.C.; FERRERES, F.; GARCIA-VIGUEIRA, C.; BANKOVA, V.S.; DE CASTRO, S.L.; DANTAS, A.P.; VALENTE, P.H.M.; PAULINO, N. Phenolic compounds from Brazilian propolis with pharmacological activities. J Ethnopharmacol. v. 74, p. 105- 112, 2001.
MARQUES-DA-SILVA, E.A.; COELHO, E.A.; GOMES, D.C.; VILELA, M.C.; MASIOLI, C.Z.; TAVARES, C.A.; FERNANDES, A.P.; AFONSO, L.C.; REZENDE, S.A. Intramuscular immunization with p36(LACK) DNA vaccine induces IFN-gamma production but does not protect BALB/c mice against Leishmania chagasi intravenous challenge.
Parasitol Res. v.98, p.67-74, 2005.
MARQUEZ, N.; SANCHO, R.; MACHO, A.; CALZADO, M.A.; FIEBICH, B.L.; MUNOZ, E. Caffeic acid phenethyl ester inhibits T-cell activation by targeting both nuclear factor of activated T-cells and NF-kappaB transcription factors. Journal of Pharmacology and
Experimental Therapeutics. v. 308, p. 993–1001, 2004.
MARTINEZ, P.A.; PETERSEN, C.A. Chronic infection by Leishmania amazonensis mediated through MAPK ERK mechanisms. Immunol Res. v. 59, p. 153–165, 2014.
MINISTÉRIO DA SAÚDE. Manual de controle da Leishmaniose Tegumentar Americana. 2ºed. Brasília/DF, 2010.
MIORIN, P.L.; LEVY JUNIOR, N.C.; CUSTODIO, A.R.; BRETZ, W.A.; MARCUCCI, M.C. Antibacterial activity of honey and propolis from Apis mellifera and Tetragonisca
angustula against Staphylococcus aureus. J Appl Microbiol. v. 95, p. 913-920, 2003.
MIRANDA, MM.; PANIS, C.; CATANEO, AHD.; DA SILVA, SS.; KAWAKAMI, NY.; LOPES, LGDF, et al. Nitric Oxide and Brazilian Propolis Combined Accelerates Tissue Repair by Modulating Cell Migration, Cytokine Production and Collagen Deposition in Experimental Leishmaniasis. Plos One. v. 10, p. 5, 2015.
MISHAN, PS.; ULRICH, N.; EPHROS, M.; ZILBERSTEIN, D. Novel intracellular SbV reducing activity correlates with antimony susceptibility in Leishmania donovani. The
Journal of Biological Chemistry. v. 276, p. 3971–3976, n. 6, 2001.
MISSIMA, F.; PAGLIARONE, A. C.; ORSATTI, C. L.; ARAÚJO JR, J. P.; SFORCIN, J. M. The Effect of Propolis on Th1/Th2 Cytokine Expression and Production by Melanomabearing Mice Submitted to Stress. Phytother Res. v. 24, p. 1501–1507, 2010.
REFERÊNCIAS
58 MOSMANN, T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Meth. v. 65, p. 55-63, 1983.
MULLANE, K.M.; R. KRAEMER et al. Myeloperoxidase activity as a quantitative assessment of neutrophil infiltration into ischemic myocardium. J. Pharmacol. Methods. v.14, n.3, p.157-67, 1985.
MURRAY, HW. Leishmaniasis in the united states: treatment in 2012. Am j trop med hyg. v. 86, n. 3, p. 340-40, 2012.
NACHER, M.; CARME, B.; SAINTE MARIE, D.; et al. Influence of clinical presentation on the efficacy of a short course of pentamidine in the treatment of cutaneous leishmaniasis in French Guiana. Ann trop med parasitol. v. 95, n. 4, p. 331-6, 2001.
NASCIMENTO, T.G. et al. Polymeric Nanoparticles of Brazilian Red Propolis Extract: Preparation, Characterization, Antioxidant and Leishmanicidal Activity. Nanoscale Research
Letters. v. 11, n. 1, p. 301, 2016.
ODA, J. M. M.; FUJITA, T.C.; PITZ, A.F., et al. Ação do extrato de própolis na Leishmaniose. Semina: Ciências Biológicas e da Saúde. v. 32, n. 1, p. 111-121, 2011.
OLIVEIRA, F.; BAFICA, A.; ROSATO, A.B.; FAVALI, C.B.F.; COSTA, J.M.; CAFÉ, V.; BARRAL-NETTO, M.; BARRAL, A. Short Report : Lesion Size Correlates with Leishmania Antigen-Stimulated TNF-Levels in Human Cutaneous Leishmaniasis. Am J Trop Med Hyg. v. 85, n. 1, p. 70–73, 2011.
OLLIARO, P.L.; GUERIN, P.J.; GERSTL, S., et al. Treatment options for visceral leishmaniasis: a systematic review of clinical studies done in India, 1980-2004. Lancet infect
dis. v. 5, n. 12, p. 763-74, 2005.
OSORIO Y FORTEA, J.; PRINA, E.; DE LA LLAVE, E.; LECOEUR, H.; LANG, T.; MILON, G. Unveiling pathways used by Leishmania amazonensis amastigotes to subvert macrophage function. Immunol. v. 219, p. 66–74, 2007.
PAULINO, N.; ABREU, S.R.L.; UTO, Y. et al. Anti-inflammatory effects of a bioavailable compound, Artepillin C, in Brazilian propolis. European Journal of Pharmacology. v. 587 p. 296–301, 2008.
PEARSON, R. D.; STEIGBIGEL, R. T. Phagocytosis and killing of the protozoan
Leishmania donovani by human polymorphonuclear leukocytes. J Immunol. v. 127(4), p. 1438-1443, 1981.
PEI, K.; OU, J.; HUANGA, J.; OUA, S. p-Coumaric acid and its conjugates: dietary sources, pharmacokinetic properties and biological activities. J Sci Food Agric. v. 96, p. 2952–2962, 2016.
PETERS, N. C.; EGEN, J. G.; SECUNDINO, N.; DEBRABANT, A.; KIMBLIN, N.; KAMHAWI, S.; LAWYER, P.; FAY, M. P.; GERMAIN, R. N.; SACKS, D. In vivo imaging reveals an essential role for neutrophils in leishmaniasis transmitted by sand flies. Science. v. 321, p. 970-973, 2008.
REFERÊNCIAS
59 PHILIPPEAUX, MM.; BARGETZI, JP.; PACHE, JC.; ROBERT, J.; SPILIOPOULOS, A.; MAUE, J. Culture and functional studies of mouse macrophages on native-like fibrillar type I collagen. European Journal of Cell Biology. v. 88. p. 243–256, 2009.
PINHEIRO, R. O.; ROSSI-BERGMANN, B. Interferon-gamma is required for the late but not early control of Leishmania amazonensis infection in C57Bl/6 mice. Memorias do
Instituto Oswaldo Cruz. v. 102, p. 79–82, 2007.
PONTIN, K.; DA SILVA FILHO, AA.; SANTOS, FF.; SILVA, MLA., et al. In vitro and in
vivo antileishmanial activities of a Brazilian green propolis extract. Parasitol Res. v. 103. p. 487–492, 2008.
REIS, L. C.; BRITO, M. E. F.; SOUZA, M. A.; PEREIRA, V. R. A. Mecanismo imunológico da resposta celular e humoral na Leishmaniose Tegumentar Americana. Rev Pat Trop. v. 35(2), p. 103-115, 2006.
ROBERTS, M.T.M. Current understandings on the immunology of leishmaniasis and recent developments in prevention and treatment. Br Med Bull. v. 75-76, n. 1, p. 115-130, 2006. SAVOIA, D. Recent updates and perspectives on leishmaniasis. J Infect Dev Ctries. v. 9, n. 6, p. 588-596, 2015.
SCOTT, P. & NOVAIS, F. O. Cutaneous leishmaniasis: immune responses in protection and pathogenesis. Nat Rev Immunol. v. 16(9), p. 581-92, 2016.
SERENO, D.; LEMESRE, J.L. Axenically cultured amastigote forms as an in vitro model for investigation of antileishmanial agents. Antimicrob. Agents Chemother., v. 41, n. 5, p. 972- 6, 1997.
SFORCIN, J.M. & BANKOVA, V. Propolis: Is there a potential for the development of new drugs? Journal of Ethnopharmacology. v. 133, p. 253–260, 2011.
SFORCIN, J.M. Biological Properties and Therapeutic Applications of Propolis. Phytother.
Res. v. 30, n. 6, p. 894-905, 2016.
SIEUWERTS, A.M.; KLIJIN, J.G.M.; PETERS, H.A.; FOEKENS, J.A. The MTT tetrazolium salt assay scrutinized: How to use this assay reliably to measure metabolic activity to cell culture in vitro for the assessment of growth characteristics, IC50- values nd cell survival. Eur Journ Clin Chem Clin Biochem, v. 33, p. 813-823, 1995.
SILVA, S. S.; THOMÉ, G. S.; CATANEO, A. H. D.; MIRANDA, M. M et al. Brazilian Propolis Antileishmanial and Immunomodulatory Effects. Evidence-Based Complementary
and Alternative Medicine. v. 2013, 2013.
SILVA, SS.; MIRANDA, MM.; COSTA, IN.; WATANABE, MAE.; PAVANELLI, WR.; FELIPE,I.; SFORCIN, JF.; COSTA, IC. Leishmanicidal activity of brazilian propolis hydroalcoholic extract in Leishmania amazonensis. Semina: Ciências Biológicas e da
Saúde. v. 36, n. 2, p. 25-34, 2015.
SOONG, L.; CHANG, C. H.; SUN, J.; LONGLEY, B. J., Jr.; RUDDLE, N. H.; FLAVELL, R. A.; MCMAHON-PRATT, D. Role of CD4+ T cells in pathogenesis associated with
REFERÊNCIAS
60
Leishmania amazonensis infection. The Journal of Immunology. v. 158, n. 11, p. 5374- 5383, 1997.
SOONG, L.; HENARD, C. A.; MELBY, P.C. Immunopathogenesis of non-healing American cutaneous leishmaniasis and progressive visceral leishmaniasis. Semin Immunopathol. v. 34, p. 735–751, 2012.
STEBUT, EV. Leishmaniasis. Journal of the German Society of Dermatology. v. 13, p. 191-200, 2015.
SUN, L.; WANG, K.; XU, X.; GE, M.; CHEN, Y.; HU, F. Potential protective effects of bioactive constituents from chinese propolis against acute oxidative stress induced by hydrogen peroxide in cardiac H9c2 cells. Evidence-Based Complementary and Alternative
Medicine.V. 2017, 2017.
SUNDAR, S.; CHAKRAVARTY, J.; RAI, V.K., et al. Amphotericin B treatment for indian visceral leishmaniasis: Response to 15 daily versus alternate-day infusions. Clin infect dis. v. 45, n. 5, p. 556-61, 2007.
SUNDAR, S.; SINGH, A. Recent developments and future prospects in the treatment of visceral leishmaniasis. Ther Adv Infectious Dis. v. 3 (3-4), p. 98-109, 2016.
SZLISZKA, E.; KUCHARSKA, A. Z.; SOKÓB-AWTOWSKA, A.; MERTAS, A.; CZUBA, Z. P.; KRÓL, W. Chemical Composition and Anti-Inflammatory Effect of Ethanolic Extract of Brazilian Green Propolis on Activated J774A.1 Macrophages. Evidence-Based
Complementary and Alternative Medicine. 2013.
TATEFUJI, T.; IZUMI, N.; OHTA, T.; ARAI, S.; IKEDA, M.; KURIMOTO, M. Isolation and identification of compounds from Brazilian propolis which enhance macrophage spreading and mobility. Biological & Pharmaceutical Bulletin. v. 19, n. 7, p. 966-970, 1996. TEIXEIRA, M.C.A.; SANTOS, RJ.; SAMPAIO, RB.; CARVALHO, LP.; SANTOS, WLC. A simple and reproducible method to obtain large numbers of axenic amastigotes of different
Leishmania species. Parasitol Res. v. 88, p. 963–968, 2002.
TITUS, R.G.; MARCHAND, M.; BOON, T.; LOUIS, J.A. A limiting dilution assay for quantifying Leishmania major in tissues of infected mice. Parasite Immunol, v.7, p.545- 555, 1985.
TRUSHEVA, B.; POPOVA, M.; BANKOVA, V.; SIMOVA, S.; MARCUCCI, MC.; MIORIN, PL.; PASIN, FR.; TSVETKOVA, I. Bioactive constituents of brazilian red propolis. Evid Based Complement Alternat Med. v. 3, p. 249-254, 2006.
UZEL, A., SORKUN, K.; ONCAG, O.; COGULU, D.; GENCAY, O.; SALIH, B. Chemical compositions and antimicrobial activities of four different Anatolian propolis samples.
Microbiol Res. v. 160, p. 189-195, 2005.
VALE, E.C.S. & FURTADO, T. Leishmaniose tegumentar no Brasil: revisão histórica da origem, expansão e etiologia. An Bras Dermatol. v. 80, n. 4, p. 421-8, 2005.
REFERÊNCIAS
61 VELASQUEZ, L.G.; GALUPPO, M.K.; REZENDE, E.; BRANDÃO, W.N.; PERON, J.P.; ULIANA,S.R.B.; DUARTE, M.I.; STOLF, B.S. Distinct courses of infection with
Leishmania (L.) amazonensis are observed in BALB/c, BALB/c nude and C57BL/6 mice.
Parasitology. v. 143, p. 692–703, 2016.
WORLD HEALTH ORGANIZATION. Leishmaniasis. Disponível em: <http://www.who.int/gho/neglected_diseases/leishmaniasis/en/>. Acesso em 05/09/2016. YAN, S.C.; LI, F.; DING, K.Y.; SUN, H. Reduction of pentavalent antimony by trypanothione and formation of a binary and ternary complex of antimony(III) and trypanothione. J. Biol. Inorg. Chem. v. 8, p. 689-697, 2003.
ZHOU, Y.; MESSIER, N.; OUELLETTE, M.; ROSEN, B.P.; MUKHOPADHYAY, R.
Leishmania major LmACR2 is a pentavalent antimony reductase that confers sensitivity to the drug Pentostam. J. Biol. Chem. v. 279, p. 37445-37451, 2004.
Certified
ISO 9001- GMP -
62 Ref. Lot: CT0216.VCOA – Certificate
of Analyzes
Product: CYTOPROPOLIS
Certified
ISO 9001- GMP -
63
Green Propolis Dried Extract at 50% w/w concentration
in Vegetal Hard Capsules of 700 mg
Botanical Origin Baccharis dracunculifolia
Certified
ISO 9001- GMP -
64 Ref. Lot: CT0216.V1.
Quality Policy
2.
Product Description
3.
Sensorial Characteristics
4.
Physic Chemical Control
5.
DPPH Free Radical Scavenging Activity
6.
Identification and Quantification of Active
Compounds
7.
Identification of the Botanical Origin
8.
Undesirable Contaminants
Certified
ISO 9001- GMP -
65
8.2
Microbiological Control
Certified
ISO 9001- GMP -
66 Ref. Lot: CT0216.V
1.
Y
Thank you very much for choosing our propolis!
We believe we have been producing the finest and
safest ftropolis on the market today. In support of
that, we have been working under the strictest
quality management system of ISO 9001, GMft &
HACCft that are issued by international
accreditation institutions.
We offer a 100% satisfaction guarantee. Despite our
efforts, should you have an unsatisfactory experience
with a particular product, simply let us know as
soon as possible, we will gladly refund your money,
or replace the merchandise.*
*
You may return items for a refund of the purchase price, or replacement for another product, within 30 days of the delivery date of your order.Certified
ISO 9001- GMP -
67 Ref. Lot: CT0216.V
Certified
ISO 9001- GMP -
68 Ref. Lot: CT0216.V
Certified
ISO 9001- GMP -
69 Ref. Lot: CT0216.V
Certified
ISO 9001- GMP -
70 Ref. Lot: CT0216.V
DESCRIPTION
Product: Propolis Dried Extract at 50% concentration in hard vegetal Capsules
Standardized Extract: Min. 50% w/w Concentration of extracted Propolis
Origin: State of Minas Gerais / Brazil
Lot code: CT0216.V
Manufactured on: February 2016
Use before: February 2019
Storage conditions: Keep the container tightly closed in a cool dry place
Ingredients: Propolis Dried Extract (from Baccharis dracunculifolia), Starch, Vegetal Capsules (V CAPS From Hypromellose)
CHARACTERISTICS
Color: Amorphous greenish powder
Appearance of the Capsules: Transparent capsules
Aroma: Characteristic of Baccharis dracunculifolia
Certified
ISO 9001- GMP -
71
PHYSIC-CHEMICAL CONTROL
Specifications Results
1 Physical Aspects Uniform, Symmetrical, Characteristic Brightness Accordingly
2 Aspect of the Content Greenish Color, No Impurities, Characteristic Odor Accordingly
3 Quercetin (Flavonoid) Min. 1450mg / 100g 2021 mg / 100g
4 Capsule moisture Max. 8% 5,90 %
5 Powder moisture Max. 8% 5,76 %
6 Phenolic Compounds
Ref. Lot: CT0216.V
4.
CONTROL
Methodology:
Min. 5000 mg /100g 7211 mg/100g 3.Japan Health Food & Nutrition Food Association (validated to October 31st, 2001).
4. ARAUJO, J.M.A. Food Chemistry, 3rd edition Instituto Adolfo Lutz; Book: Rules of the Instituto Adolfo Lutz, v.1: Physical and Chemical Methods for Analysis of Food. 1ed. Digital. São Paulo: 2005.p.98.
5. ARAUJO, J.M.A. Food Chemistry, 3rd edition Instituto Adolfo Lutz; Book: Rules of the Instituto Adolfo Lutz, v.1: