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Avalia??o da atividade citot?xica in vitro dos extratos vegetais de Pseudobrickellia brasiliensis (Spreng) R. M. King & H. Rob, Miconia ferruginata DC e Ageratum fastigiatum (Gardn.) R. M. King sobre c?lulas tumorais JurkatC?rdoba, Mar?a Ang?lica Mera 15 March 2017 (has links)
?rea de concentra??o: Ci?ncias farmac?uticas. / Incluir como ag?ncias financiadoras: Grupo Coimbra das Universidades Brasileiras (GCUB) e Organiza??o
dos Estados Americanos (OEA). / Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2018-01-03T17:55:46Z
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Previous issue date: 2017 / O Cerrado mineiro possui muitas esp?cies vegetais utilizadas na medicina
popular, dentre elas est?o a Pseudobrickellia brasilensis , a Miconia ferruginata e
Ageratum fastigiatum, plantas popularmente usadas como analg?sico, cicatrizante e
anti-inflamat?rio. Estudos realizados no Laborat?rio de Imunologia da UFVJM
indicaram que extratos destas plantas possuem efeito inibit?rio sobre a ativa??o de
linf?citos humanos in vitro. Em virtude disso, e com base em relatos populares sobre
uma poss?vel a??o antitumoral, o objetivo principal deste trabalho foi pesquisar se estes
extratos apresentariam ou n?o efeito citot?xico sobre a linhagem tumoral Jurkat,
utilizando-se concentra??es n?o t?xicas, j? avaliadas sobre c?lulas mononucleares do
sangue perif?rico humano (PBMC). Uma vez que os extratos vegetais foram dilu?dos no
solvente Dimetilsulf?xido (DMSO), inicialmente investigou-se a concentra??o m?xima
de DMSO que n?o alteraria a viabilidade das c?lulas Jurkat ap?s 24, 48 e 72 horas de
cultura. O efeito citot?xico dos extratos foi avaliado sobre linf?citos tumorais (Jurkat) e
sobre linf?citos humanos de volunt?rios h?gidos pelo m?todo de exclus?o com azul de
tripan, com exce??o das culturas tratadas com P.brasilensis, onde a viabilidade foi
avaliada por citometria de fluxo. Tamb?m foram calculados os ?ndices de seletividade e
percentuais de efic?cia dos extratos com rela??o ao f?rmaco antineopl?sico Paclitaxel.
Foi avaliada tamb?m a interfer?ncia desses extratos sobre as fases do ciclo celular das
c?lulas Jurkat, bem como os mecanismos de morte envolvidos na a??o citot?xica dos
extratos. De acordo com os resultados obtidos, o DMSO n?o apresentou efeito t?xico
sobre as c?lulas Jurkat na concentra??o de 1%v/v nos tr?s tempos de incuba??o, sendo
esta a concentra??o de solvente utilizada em todos os ensaios realizados. Extratos
etan?licos das folhas da P. brasilensis (PBf) a 200?g/mL, das partes a?reas da A.
fastigiatum (Afpa) a 50?g/mL e extratos etan?licos e aquosos da M. ferruginata (Mfet,
Mfaq) a 125?g/mL mostraram maior toxicidade sobre as c?lulas Jurkat principalmente
ap?s 72h de tratamento. O tratamento, por 24h, com extrato Afpa 50 ?g/mL mostrou ser
o mais seletivo e eficaz com rela??o aos outros extratos. Foi evidenciado em todos os
extratos, que nas maiores concentra??es, ap?s de 24 horas de tratamento, houve uma
inibi??o na progress?o da fase G2M do ciclo celular. O extrato das partes a?reas de
Ageratum fastigiatum tamb?m produziu uma reten??o dessas c?lulas na fase G0-G1.
Todos os extratos induziram a apoptose as c?lulas Jurkat, onde o n?mero de c?lulas em
apoptose tardio foi predominante em rela??o aos processos necr?ticos. / Disserta??o (Mestrado) ? Programa de P?s-gradua??o em Ci?ncias Farmac?uticas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2017. / The Cerrado Mineiro has many vegetable species used in folk medicine. Plants like
Pseudobrickellia Brasilensis, Miconia ferruginata and Ageratum fastigiatum are widely
used as analgesic, healing or anti-inflammatory. Studies carried on the Immunology
Laboratory (UFVJM) shown that extracts from these plants has inhibitory effects in the
activation of in-vitro-cultured human lymphocytes. In this way, and based on folk
stories related with their anti-tumor action, the main objective of this work was research
about the cytotoxic effect of this plants on Jurkat tumor line-cell cultures, using as first
experiment non-toxic extract concentrations previously proved on peripheral blood
mononuclear cells (PBCM). Once the vegetable extracts were diluted in dimetilsulfoxide
(DMSO), previously, the maximum DMSO concentration not-altering cell
viability was determined after 24, 48 and 72 hours. Cytotoxic effect was studied using
the exclusion method with tripam blue on tumor lymphocytes (Jukart) and human
lymphocytes, donated by healthy volunteers, except for P.brasilensis treated line cells,
for which cell viability was studied by flux cytometer. The selectivity index and extracts
efficiency percentage were calculated and related with the antineoplastic Paclitaxel
drug. The extracts interference on the Jukart cell cycle phases and the cell death
mechanisms related with the cytotoxic activity were evaluated. According to the
obtained results, DMSO does not show cytotoxic effects on Jukart cells in 1% (v/v)
concentration at the three incubation times, being this the solvent concentration used in
all the experiments. Ethanol extracts of P. brasilensis leaves (PBf) at 200?g / mL, aerial
parts of A. fastigiatum (Afpa) at 50?g / mL and M. ferruginata aqueous and ethanolic
extracts (Mfet, Mfaq) at 125?g / mL showed higher Toxicity on Jurkat cells mainly
after 72h of treatment. The 24 h treatment with Afpa at 50 ?g/mL was the most
selective and effective in comparison with the other tested extracts. It was evidenced in
all extracts, that at the highest concentrations, after 24 hours of treatment, there was an
inhibition in the progression of the G2M phase of the cell cycle. The extract of
Ageratum fastigiatum aerial parts also produced the G0-G1 phase retention. All extracts
induced Jukart cell apoptosis, and cell number on late apoptotic phase predominated on
the necrotic ones.
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