Made available in DSpace on 2014-06-11T19:33:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-12-11Bitstream added on 2014-06-13T20:24:56Z : No. of bitstreams: 1 scolastici_c_dr_botfm.pdf: 429755 bytes, checksum: 699c635f6217a38ea4f5d62a997ee8ce (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O licopeno e um pigmento natural sintetizado por plantas e microorganismos, e encontrado principalmente no tomate. E um isomero aciclico do À-caroteno sem atividade pro-vitaminica-A, sendo um dos mais potentes agentes antioxidantes. Diversos pesquisadores tem demonstrado o efeito protetor do licopeno contra danos no DNA e sobre a carcinogenese quimicamente induzida, embora os mecanismos envolvidos nesses processos nao estejam, ainda, totalmente esclarecidos. Assim, o presente estudo objetivou avaliar o potencial quimioprotetor do licopeno contra danos no DNA e sobre a hepaticarcinogenese em roedores. O efeito anticarcinogenico do licopeno em figado de ratos foi avaliado utilizando-se os focos GST-P positivos (modelo de hepatocarcinogenese de media duracao proposto por Ito et al., 1988) como biomarcadores de lesoes pre-neoplasicas. Para avaliacao do efeito antigenotoxico e antimutagenico do carotenoide, bem como de seu possivel mecanismo de acao, foram utilizados, respectivamente, o teste do cometa e o teste do micronucleo em duas linhagens celulares in vitro: celulas de ovario de hamster chines (CHO) e celulas de hepatoma humano (HepG2). Mutagenos de acao direta (peroxido de hidrogenio, oxido de 4-nitroquinolina - 4NQO e metil metanosulfonato - MMS) e indireta (dietilnitrosamina -DEN), foram utilizados como indutores de danos no DNA in vitro. Os resultados mostraram que o licopeno, na concentracao 300 ppm, reduziu significativamente os danos no DNA induzidos pela DEN na etapa de iniciacao da hepatocarcinogenese de rato, embora nao tenha sido observada reducao do numero e area dos focos GST-P-positivos. Os resultados dos experimentos in vitro mostraram que o tratamento previo e simultaneo com licopeno (10, 25, e 50 ÊM) foi eficaz em reduzir os niveis de danos no DNA induzidos pelo H2O2 e pela DEN em celulas HepG2, quando avaliados tanto teste do cometa como do micronucleo. / Lycopene is a natural pigment synthesized by plants and microorganisms, and mainly found in tomatoes. It is an acyclic isomer of â-carotene with no vitamin A activity, and one of the most potent antioxidants. Several studies have showed the chemopreventive effect of lycopene on chemically-induced DNA damage and on chemical hepatocarcinogenesis, however its mechanism of action needs to be clarified. The present study aimed to evaluate the protective activity of lycopene on chemically-induced DNA damage and on rodent hepatocarcinogenesis. Anticarcinogenic potential of lycopene was evaluated using GST-P positive foci (medium-term hepatocarcinogenesis model described by Ito et al., 1988) as a biomarker of preneoplasic lesion. Lycopene antigenotoxicity and antimutagenicity, as well as its mechanism of action were investigated in two cell lines (CHO and human hepatoma cell HepG2) using the comet assay and micronucleus test, respectively. Direct-acting (hydrogen peroxide; 4-nitroquinoline 1-oxide - 4NQO; methyl methanesulphonate - MMS) and indirect-acting (n-nitrosodiethylamine DEN) mutagens were used to induce in vitro DNA damage. The results showed that lycopene (300 ppm) reduced DEN-induced DNA damage at the initiation step of in vivo hepatocarcinogenesis, although no effect was observed on the number and area of GST-P positive foci in liver of rats. Data from in vitro experiments showed that lycopene (10, 25 and 50 ìM) was effective in reducing DNA damage induced by H2O2 and DEN in HepG2 cells, both in comet assay and in micronucleus test. In CHO cells the chemopreventive acitivity of lycopene was visualized only for primary DNA damage in the comet assay. In conclusion our data confirmed the chemopreventive effect of lycopene on chemically-induced DNA damage, although such activity depends on the treatment schedule used. On the other hand, no protective action of this carotenoid on DEN-induced preneoplasic lesion in liver of rats was detected.

Resumo: O licopeno e um pigmento natural sintetizado por plantas e microorganismos, e encontrado principalmente no tomate. E um isomero aciclico do À-caroteno sem atividade pro-vitaminica-A, sendo um dos mais potentes agentes antioxidantes. Diversos pesquisadores tem demonstrado o efeito protetor do licopeno contra danos no DNA e sobre a carcinogenese quimicamente induzida, embora os mecanismos envolvidos nesses processos nao estejam, ainda, totalmente esclarecidos. Assim, o presente estudo objetivou avaliar o potencial quimioprotetor do licopeno contra danos no DNA e sobre a hepaticarcinogenese em roedores. O efeito anticarcinogenico do licopeno em figado de ratos foi avaliado utilizando-se os focos GST-P positivos (modelo de hepatocarcinogenese de media duracao proposto por Ito et al., 1988) como biomarcadores de lesoes pre-neoplasicas. Para avaliacao do efeito antigenotoxico e antimutagenico do carotenoide, bem como de seu possivel mecanismo de acao, foram utilizados, respectivamente, o teste do cometa e o teste do micronucleo em duas linhagens celulares in vitro: celulas de ovario de hamster chines (CHO) e celulas de hepatoma humano (HepG2). Mutagenos de acao direta (peroxido de hidrogenio, oxido de 4-nitroquinolina - 4NQO e metil metanosulfonato - MMS) e indireta (dietilnitrosamina -DEN), foram utilizados como indutores de danos no DNA in vitro. Os resultados mostraram que o licopeno, na concentracao 300 ppm, reduziu significativamente os danos no DNA induzidos pela DEN na etapa de iniciacao da hepatocarcinogenese de rato, embora nao tenha sido observada reducao do numero e area dos focos GST-P-positivos. Os resultados dos experimentos in vitro mostraram que o tratamento previo e simultaneo com licopeno (10, 25, e 50 ÊM) foi eficaz em reduzir os niveis de danos no DNA induzidos pelo H2O2 e pela DEN em celulas HepG2, quando avaliados tanto teste do cometa como do micronucleo / Abstract: Lycopene is a natural pigment synthesized by plants and microorganisms, and mainly found in tomatoes. It is an acyclic isomer of â-carotene with no vitamin A activity, and one of the most potent antioxidants. Several studies have showed the chemopreventive effect of lycopene on chemically-induced DNA damage and on chemical hepatocarcinogenesis, however its mechanism of action needs to be clarified. The present study aimed to evaluate the protective activity of lycopene on chemically-induced DNA damage and on rodent hepatocarcinogenesis. Anticarcinogenic potential of lycopene was evaluated using GST-P positive foci (medium-term hepatocarcinogenesis model described by Ito et al., 1988) as a biomarker of preneoplasic lesion. Lycopene antigenotoxicity and antimutagenicity, as well as its mechanism of action were investigated in two cell lines (CHO and human hepatoma cell HepG2) using the comet assay and micronucleus test, respectively. Direct-acting (hydrogen peroxide; 4-nitroquinoline 1-oxide - 4NQO; methyl methanesulphonate - MMS) and indirect-acting (n-nitrosodiethylamine DEN) mutagens were used to induce in vitro DNA damage. The results showed that lycopene (300 ppm) reduced DEN-induced DNA damage at the initiation step of in vivo hepatocarcinogenesis, although no effect was observed on the number and area of GST-P positive foci in liver of rats. Data from in vitro experiments showed that lycopene (10, 25 and 50 ìM) was effective in reducing DNA damage induced by H2O2 and DEN in HepG2 cells, both in comet assay and in micronucleus test. In CHO cells the chemopreventive acitivity of lycopene was visualized only for primary DNA damage in the comet assay. In conclusion our data confirmed the chemopreventive effect of lycopene on chemically-induced DNA damage, although such activity depends on the treatment schedule used. On the other hand, no protective action of this carotenoid on DEN-induced preneoplasic lesion in liver of rats was detected / Orientador: Daisy Maria Fávero Salvatori / Coorientador: Luís Fernando Barbisan / Banca: Thomas Prates Ong / Banca: Maria Lúcia Zaidan Dagli / Banca: Eliana Varanda / Banca: Ana Lúcia dos Anjos Ferreira / Doutor