Effects of ethanolic, butanolic or aqueous extracts of Pfaffia paniculata on human breast tumor cell lines proliferation

Dissertacao de Mestrado Autor Nagamine, Marcia Kazumi (Catalogo USP) Titulo em portugues Efeitos dos extratos etanolico, butanolico ou aquoso de Pfaffia paniculata sobre a proliferacao de linhagens tumorais de celulas mamarias humanas Palavras-chave em portugues Cultura de celulas Neoplasias Plantas medicinais Resumo em portugues As raizes de Pfaffia paniculata (Ginseng brasileiro) sao comercialmente encontradas como capsulas contendo as raizes pulverizadas, misturadas ou nao ao extrato etanolico destas raizes. Estas raizes sao popularmente recomendadas para varios propositos, e tem sido utilizadas na terapia contra o câncer pela medicina popular. Os principais componentes encontrados nestas raizes ja isolados incluem o acido pfaffico e os pfaffosideos A, C, D, E e F; estes componentes inibiram o crescimento de celulas do melanoma B-16, demonstrado em um outro estudo. O objetivo deste estudo foi investigar os efeitos dos extratos etanolico, butanolico ou aquoso das raizes de Pfaffia paniculata sobre o crescimento das linhagens tumorais de celulas mamarias humanas MCF-7 e SKBR-3, utilizando metodo colorimetrico (cristal violeta) e quantificacao das celulas que incorporaram bromodeoxiuridina (BrdU). A coloracao por laranja de acridina/brometo de etideo foi utilizada para avaliar morte celular, e as alteracoes subcelulares foram avaliadas por microscopia eletronica. O extrato butanolico apresentou efeitos citotoxicos nas linhagens MCF-7 e SKBR-3. Morte celular foi observada pelo tratamento com o extrato butanolico por 1 h; alteracoes morfologicas foram observadas com 500μg/mL deste extrato, apos 24 h de tratamento. Apos o tratamento por 48 h com o extrato butanolico nesta mesma concentracao, foi observado degeneracao de componentes citoplasmaticos e alteracoes morfologicas sugestivas de morte celular. O tratamento com 1000μg/mL deste extrato levou a profundas alteracoes citoplasmaticas e nucleares, incompativeis com a viabilidade celular. O tratamento com o extrato etanolico nao causou efeitos significantes no crescimento das celulas MCF-7 e SKBR-3; o extrato aquoso, por outro lado, estimulou o crescimento das celulas MCF-7, apos o tratamento por 1 h. Estes resultados indicam efeitos citotoxicos exercidos pelo extrato butanolico das raizes de Pfaffia paniculata em linhagens celulares de tumor de mama humano Titulo em ingles Effects of ethanolic, butanolic or aqueous extracts of Pfaffia paniculata on human breast tumor cell lines proliferation Palavras-chave em ingles Cell culture Medicinal plants Neoplasia Resumo em ingles The roots of Pfaffia paniculata (Brazilian ginseng) are commercially available as capsules containing the powdered roots, mixed or not with the ethanolic extract of the roots. These roots have been populary recommended for many purposes, and have also been used on cancer therapy by folk medicine. The main components found in these roots that have already been isolated include pfaffic acid and pfaffosides A, C, D, E and F; these components inhibited the growth of melanoma B-16 cells, as shown in another study. The aim of this study was to investigate the effects of ethanolic, butanolic or aqueous extract of the roots of Pfaffia paniculata on the growth of human breast tumor cell lines MCF-7 and SKBR-3, using a colorimetric method (crystal violet) and quantification of bromodeoxiuridine (BrdU) positive cells. Acridine orange/ethidium bromide staining was utilized to evaluate cell death, and subcellular alteration was evaluated by electron microscopy. The butanolic extract presented cytotoxic effects in MCF-7 and SKBR-3 cell lines. Cell death was observed following treatment with the butanolic extract for 1 h; morphologic alterations were observed with 500μg/mL of this extract, after 24 and 48 h of treatment. Following treatment for 48 h with the butanolic extract with this same concentration, degeneration of cytoplasmic components and morphological alterations suggestive of cell death were observed. Treatment with 1000μg/mL of this extract lead to profound cytoplasmic and nuclear alterations, incompatible with cell viability. The treatment with ethanolic extract didn t lead to significant effects on the growth of MCF-7 and SKBR-3 cells; the aqueous extract, on the other hand, stimulated the growth of MCF-7 cells, following treatment for 1 hour. These results point to cytotoxic effects exerted by the butanolic extract from the roots of Pfaffia paniculata on human breast tumor cell lines