Efeito de extrato de Cytinus spp num modelo animal K14- HPV16
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2022-04-29
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As plantas medicinais têm contribuído fortemente para o desenvolvimento de formulações/produtos com potencial terapêutico porque muitos dos seus extratos possuem propriedades anti-inflamatórias, antimicrobianas e antitumorais. Os extratos de Cytinus hypocistis possuem substâncias com propriedades antioxidante, anti-inflamatória, antibacteriana e antiproliferativa, sendo promissoras do ponto de vista terapêutico. As infeções por certos tipos de Vírus do Papiloma Humano (HPV), como o HPV16, estão associadas ao desenvolvimento de vários cancros humanos, incluindo o cancro do colo do útero, neoplasias anogenitais e um número crescente de cancros de cabeça e pescoço; deste modo, o HPV constitui um grande risco para a Saúde Pública. Pretendemos com este trabalho estudar a influência da aplicação tópica de um extrato obtido a partir de Cytinus hypocistis (L.) L. sobre parâmetros fisiológicos do murganho K14-HPV16 com o objetivo de compreender o seu efeito terapêutico nas lesões espontâneas desenvolvidas neste modelo animal. Para a realização deste estudo foram usados trinta murganhos fêmeas, da estirpe FVB/n, com idades compreendidas entre as 33-37 semanas, sendo 10 wild-type e 20 transgénicas: Grupo 1 (K14-HPV16, n=5) tratados com concentração 1 do extrato; grupo 2 (K14-HPV16, n=5) tratados com a concentração 2 do extrato; grupo 3 (K14-HPV16, n=5) tratados com a concentração 3 do extrato; grupo 4 (wild-type, n=5) tratados com a concentração 3 do extrato; os grupos 5 (K14-HPV16, n=5) e 6 (wild-type, n=5) foram usados como controlo. O extrato foi aplicado por via tópica no pavilhão auricular dos animais, uma vez por dia, ao longo de 28 dias, com exceção dos fins de semana, com auxílio de uma espátula. No final do ensaio os animais foram eutanasiados pela administração de uma overdose da associação de xilazina com ketamina, sendo realizada uma necrópsia completa. Os órgãos dos animais foram recolhidos, registada a sua massa e feita a sua análise histológica. O sangue foi recolhido por punção cardíaca direta para análise bioquímica, o microhematócrito também foi realizado. Verificou-se que a aplicação do extrato mostrou não ter influência na ingestão média de alimento (consumo final (g): G1-4,39; G2-4,10; G3-3,86; G-3,57; G5-4,23; G6-3,15) e de água (consumo final (g): G1-8,62; G2-8,82; G3-8,05; G4-4,7; G5-8,39; G6-3,99), na massa dos órgãos (media fígado (g): G1-1,496±0,042; G2-1,550±0,083; G3-1,424±0,092; G4- 1,550±0,118; G5-1,40±0,083; G6-1,238±0,062), no microhematócrito, no parâmetro de stresse XXII oxidativo e nos parâmetros bioquímicos. A histologia demonstrou a presença de lesões proliferativas na pele e na mucosa oral dos animais K14-HPV16, sem associação com a aplicação do extrato; foi observada a presença de infiltrado inflamatório nos órgãos internos de todos os grupos; os animais K14-HPV16 apresentaram uma diminuição na média de massa corporal ((g): G1-27,30±0,45; G2-24,69±0,66; G3-24,20±0,89; G4-30,08±1,69; G5- 25,54±0,93; G6-27,02±0,75), independentemente da concentração de extrato aplicado. Em conclusão, os resultados indicam que a aplicação do extrato de Cytinus hypocistis não teve influências nas lesões provocadas pelo HPV, sendo as diferenças observadas associadas ao fenótipo dos animais. No geral foi bem tolerado pelos animais, contudo estudos adicionais são necessários para explicitar o potencial genotóxico do extrato e a segurança da sua utilização.
Medicinal plants have strongly contributed to the development of formulations/products with therapeutic potential because many of their extracts have anti-inflammatory, antimicrobial and antitumor properties. Cytinus hypocistis extracts have substances with antioxidant, antiinflammatory, antibacterial and antiproliferative properties, promising from a therapeutic point of view. Infections with certain types of Human Papilloma Virus (HPV), such as HPV16, are associated with the development of several human cancers, including cervical cancer, anogenital neoplasms and an increasing number of head and neck cancers; thus, HPV poses a major public health risk. With this work, we intend to study the influence of the topical application of an extract obtained from Cytinus hypocistis (L.) L. on physiological parameters of the K14-HPV16 mouse in order to understand its therapeutic effect on the spontaneous lesions developed in this animal model. We have used 30 female mice of the FVB/n strain, with ages between 33-37 weeks, including 10 wild-type and 20 transgenic. The group 1 (K14-HPV16, n=5) was treated with concentration 1 of the extract; group 2 (K14-HPV16, n=5) was treated with concentration 2 of the extract; group 3 (K14-HPV16, n=5) treated with concentration 3 of the extract; group 4 (wild-type, n=5) treated with concentration 3 of the extract; group 5 (K14-HPV16, n=5) and group 6 (wild-type, n=5) were used as controls. The extract was applied topically on the ear of the animals, once a day, for 28 days, except for weekends, with the aid of a spatula. At the end of experiment, the mice were euthanized with xylazine and ketamine and a complete necropsy was performed. The organs were collected and their weight was recorded, being fixed with formaldehyde for subsequent histological analysis. The blood was extracted through direct cardiac puncture for biochemical and microhematocrit analysis. It was found that the application of the extract proved to have no influence on average food intake (final intake (g): G1-4.39; G2-4.10; G3-3.86; G-3.57; G5-4.23; G6-3.15) and water intake (final intake (g): G1-8.62; G2-8.82; G3-8.05; G4-4.7; G5-8.39; G6-3.99), in organ mass (mean liver (g): G1-1.496±0.042; G2-1.550±0.083; G3-1.424±0.092; G4-1.550±0.118; G5- 1.40±0.083; G6-1.238±0.062), in microhematocrit, oxidative stress parameter and biochemical parameters. Histology demonstrated the presence of proliferative epithelial lesions in skin and XXIV oral mucosa of K14-HPV16 animals, with no association with the application of the extract. Inflammatory focal infiltrate was observed in internal organs of all groups. K14-HPV16 animals showed a decrease in mean body mass (g): G1-27.30±0.45; G2-24.69±0.66; G3- 24.20±0.89; G4-30.08±1.69; G5-25.54±0.93; G6-27.02±0.75), regardless of the concentration of extract applied. In conclusion, the results indicate that the application of Cytinus hypocistis extract had no influence on the lesions caused by HPV, with the observed differences associated with the phenotype of the animals. In general, it was well tolerated by the animals; however, further studies are needed to explain the genotoxic potential of the extract and the safety of its use.
Medicinal plants have strongly contributed to the development of formulations/products with therapeutic potential because many of their extracts have anti-inflammatory, antimicrobial and antitumor properties. Cytinus hypocistis extracts have substances with antioxidant, antiinflammatory, antibacterial and antiproliferative properties, promising from a therapeutic point of view. Infections with certain types of Human Papilloma Virus (HPV), such as HPV16, are associated with the development of several human cancers, including cervical cancer, anogenital neoplasms and an increasing number of head and neck cancers; thus, HPV poses a major public health risk. With this work, we intend to study the influence of the topical application of an extract obtained from Cytinus hypocistis (L.) L. on physiological parameters of the K14-HPV16 mouse in order to understand its therapeutic effect on the spontaneous lesions developed in this animal model. We have used 30 female mice of the FVB/n strain, with ages between 33-37 weeks, including 10 wild-type and 20 transgenic. The group 1 (K14-HPV16, n=5) was treated with concentration 1 of the extract; group 2 (K14-HPV16, n=5) was treated with concentration 2 of the extract; group 3 (K14-HPV16, n=5) treated with concentration 3 of the extract; group 4 (wild-type, n=5) treated with concentration 3 of the extract; group 5 (K14-HPV16, n=5) and group 6 (wild-type, n=5) were used as controls. The extract was applied topically on the ear of the animals, once a day, for 28 days, except for weekends, with the aid of a spatula. At the end of experiment, the mice were euthanized with xylazine and ketamine and a complete necropsy was performed. The organs were collected and their weight was recorded, being fixed with formaldehyde for subsequent histological analysis. The blood was extracted through direct cardiac puncture for biochemical and microhematocrit analysis. It was found that the application of the extract proved to have no influence on average food intake (final intake (g): G1-4.39; G2-4.10; G3-3.86; G-3.57; G5-4.23; G6-3.15) and water intake (final intake (g): G1-8.62; G2-8.82; G3-8.05; G4-4.7; G5-8.39; G6-3.99), in organ mass (mean liver (g): G1-1.496±0.042; G2-1.550±0.083; G3-1.424±0.092; G4-1.550±0.118; G5- 1.40±0.083; G6-1.238±0.062), in microhematocrit, oxidative stress parameter and biochemical parameters. Histology demonstrated the presence of proliferative epithelial lesions in skin and XXIV oral mucosa of K14-HPV16 animals, with no association with the application of the extract. Inflammatory focal infiltrate was observed in internal organs of all groups. K14-HPV16 animals showed a decrease in mean body mass (g): G1-27.30±0.45; G2-24.69±0.66; G3- 24.20±0.89; G4-30.08±1.69; G5-25.54±0.93; G6-27.02±0.75), regardless of the concentration of extract applied. In conclusion, the results indicate that the application of Cytinus hypocistis extract had no influence on the lesions caused by HPV, with the observed differences associated with the phenotype of the animals. In general, it was well tolerated by the animals; however, further studies are needed to explain the genotoxic potential of the extract and the safety of its use.
Descrição
Dissertação de Mestrado em Biologia Clínica Laboratorial
Palavras-chave
Animal , In vivo