Healing activity assessement of the ethanolic extract and Triterpene 3Β, 6 Β, 16 Β Trihydroxylup-20 (29) â Eno from combretum leprosum leafs in the treatment of cutaneos lesions induced in mice

Detalhes bibliográficos
Autor(a) principal: LÃvia Coelho de Assis
Data de Publicação: 2015
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFC
Texto Completo: http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=15556
Resumo: Wound healing is a complex and dynamic process involving an integrated action of several cell types, extracellular matrix and soluble mediators such as cytokines, which serve to recover the integrity of damaged anatomical structure. Natural products are the basis for many biotechnological inputs, as they are biologically active compounds with great potential for therapeutics and can provide molecular basis for many drugs in the clinics. Combretum leprosum Mart. is a native species of Caatinga, important biome in the Brazilian semiarid region, being popularly used in Brazil for the treatment of many diseases such as skin diseases. This study aimed to investigate the pro-healing potential of the ethanol extract (EECL) and the bioactive triterpene 3&#946;, 6 &#946;, 16 &#946;-trihydroxylup-20 (29) ene (CLF-1) isolated from leaves of C. leprosum. Cell migration assay was performed using fibroblasts, which were treated with CLF-1 (1.75 Âg / ml and 2.5 Âg / ml) and analyzed after 0, 6 and 24 hours. Skin wounds (1 cm2) were induced in the dorsal region of mice (n = 30) and were treated with daily topical application (100Âl) according to the group: saline, EECL (100&#956;g / ml) and CLF-1 (100&#956;g / ml) for 12 days. The results showed that CLF-1 did not show toxicity to fibroblasts and further increased cell proliferation and migration so that a significant increase on injury closing percentage was observed using both concentrations (1.75 mg / ml vs 2.5 mg / mL,) compared to control (p <0.05). Within 24 hours, there was also a significant increase in the rate of migration comparing CLF-1 at 2.5 ug / ml and control group (p<0.05). For the in vivo analysis, on the 2nd post surgical day (PS), the lesions treated with EECL and CLF-1 showed moderate presence of vessels in granulation tissue progressing to the dermis, as well as a slight inflammatory infiltrate. It was also observed that, on the 7th PS day, the animals treated with CLF-1 showed moderate angiogenesis, whereas animals treated with saline showed cell proliferation and migration into the wound. Furthermore, on the 12th day PS, the animals treated with EECL and CLF-1 showed complete closing of the injury, while those receiving saline exhibited open wounds. Thus, it was concluded that EECL and CLF-1 showed pro-healing effects, as well as a possible anti-inflammatory action and additional research is needed to elucidate its mechanisms in the wound healing process.
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spelling info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisHealing activity assessement of the ethanolic extract and Triterpene 3&#914;, 6 &#914;, 16 &#914; Trihydroxylup-20 (29) â Eno from combretum leprosum leafs in the treatment of cutaneos lesions induced in miceAvaliaÃÃo da atividade cicatrizante do extrato etanÃlico e do Triterpeno 3, 6 , 16 Trihydroxylup-20 (29) - Eno provenientes de folhas de Combretum leprosum mart. no tratamento de lesÃes cutÃneas induzidas em camundongos2015-08-28Edson Holanda Teixeira71755985304http://lattes.cnpq.br/8800103074322967 Max Victor Carioca Freitas67953662353http://lattes.cnpq.br/2609171032311880Maria Jania Teixeira23305908300http://buscatextual.cnpq.br/buscatextual/visualizacv.jsp?id=K4798130Z1Helcio Silva dos Santos77045050330http://lattes.cnpq.br/347884205498449702898042307http://lattes.cnpq.br/7104770080836632LÃvia Coelho de AssisUniversidade Federal do CearÃPrograma de PÃs-GraduaÃÃo em PatologiaUFCBRANATOMIA PATOLOGICA E PATOLOGIA CLINICAWound healing is a complex and dynamic process involving an integrated action of several cell types, extracellular matrix and soluble mediators such as cytokines, which serve to recover the integrity of damaged anatomical structure. Natural products are the basis for many biotechnological inputs, as they are biologically active compounds with great potential for therapeutics and can provide molecular basis for many drugs in the clinics. Combretum leprosum Mart. is a native species of Caatinga, important biome in the Brazilian semiarid region, being popularly used in Brazil for the treatment of many diseases such as skin diseases. This study aimed to investigate the pro-healing potential of the ethanol extract (EECL) and the bioactive triterpene 3&#946;, 6 &#946;, 16 &#946;-trihydroxylup-20 (29) ene (CLF-1) isolated from leaves of C. leprosum. Cell migration assay was performed using fibroblasts, which were treated with CLF-1 (1.75 Âg / ml and 2.5 Âg / ml) and analyzed after 0, 6 and 24 hours. Skin wounds (1 cm2) were induced in the dorsal region of mice (n = 30) and were treated with daily topical application (100Âl) according to the group: saline, EECL (100&#956;g / ml) and CLF-1 (100&#956;g / ml) for 12 days. The results showed that CLF-1 did not show toxicity to fibroblasts and further increased cell proliferation and migration so that a significant increase on injury closing percentage was observed using both concentrations (1.75 mg / ml vs 2.5 mg / mL,) compared to control (p <0.05). Within 24 hours, there was also a significant increase in the rate of migration comparing CLF-1 at 2.5 ug / ml and control group (p<0.05). For the in vivo analysis, on the 2nd post surgical day (PS), the lesions treated with EECL and CLF-1 showed moderate presence of vessels in granulation tissue progressing to the dermis, as well as a slight inflammatory infiltrate. It was also observed that, on the 7th PS day, the animals treated with CLF-1 showed moderate angiogenesis, whereas animals treated with saline showed cell proliferation and migration into the wound. Furthermore, on the 12th day PS, the animals treated with EECL and CLF-1 showed complete closing of the injury, while those receiving saline exhibited open wounds. Thus, it was concluded that EECL and CLF-1 showed pro-healing effects, as well as a possible anti-inflammatory action and additional research is needed to elucidate its mechanisms in the wound healing process.A cicatrizaÃÃo cutÃnea à um processo complexo e dinÃmico que envolve uma aÃÃo integrada de vÃrios tipos celulares, matriz extracelular e mediadores solÃveis como citocinas, que tem por finalidade a recuperaÃÃo da integridade da estrutura anatÃmica danificada. Produtos naturais sÃo a base para diversos insumos biotecnolÃgicos, pois sÃo compostos biologicamente ativos com grande potencial terapÃutico, e que podem formar bases moleculares para muitas drogas em uso clÃnico. Combretum leprosum Mart. à uma espÃcie nativa da Caatinga, bioma importante na regiÃo do semiÃrido brasileiro, sendo esta espÃcie popularmente utilizada no Brasil para o tratamento de diversas doenÃas, tais como doenÃas de pele. O presente estudo investigou o potencial prÃ-cicatrizante do extrato etanÃlico (EECL) e do triterpeno bioativo 3&#946;, 6 &#946;, 16 &#946;-trihydroxylup-20 (29)-eno (CLF-1) isolado das folhas de C. leprosum. Para isto, o ensaio de migraÃÃo de cÃlulas foi realizado com fibroblastos, que foram pÃs tratados com CLF-1 (1,75 &#956;g/mL e 2,5 &#956;g/mL) e analisados em 0, 6 e 24 horas. Feridas cutÃneas (1 cm2) foram realizadas na zona dorsal de camundongos (n=30) e em seguida foram tratadas com uma aplicaÃÃo tÃpica diÃria (100 &#956;l) de acordo com o grupo: salina, EECL (100&#956;g/mL) e CLF-1 (100&#956;g/mL) durante 12 dias. Os resultados demonstraram que CLF-1 nÃo apresentou toxicidade aos fibroblastos e, ainda, em 6 horas estimulou a proliferaÃÃo e migraÃÃo celular, de modo que observou-se um aumento significativo na porcentagem de fechamento em ambas concentraÃÃes (1,75 &#956;g/mL e 2,5 &#956;g/mL) em relaÃÃo ao controle (p<0,05). Em 24 horas, verificou-se tambÃm aumento significativo na taxa fechamento quando comparado CLF-1 2,5 &#956;g/mL em relaÃÃo ao controle (p<0,05). Na avaliaÃÃo in vivo, observou-se no 2 dia pÃs cirÃrgico (P.C.), que as lesÃes tratadas com EECL e CLF-1 mostraram presenÃa moderada de vasos no tecido de granulaÃÃo, progredindo na derme, bem como apresentaram um leve infiltrado inflamatÃrio. Observou-se, ainda, que no 7 dia P.C. os animais tratados com CLF-1 demonstraram moderada angiogÃnese, enquanto que os animais tratados com salina apresentaram proliferaÃÃo e migraÃÃo celular em direÃÃo ao leito da ferida. Ademais, no 12 dia P.C., os animais tratados com EECL e CLF-1 apresentaram o fechamento completo da lesÃo, enquanto os que receberam salina, apresentaram, ainda, feridas abertas. Desta forma, foi possÃvel concluir que o EECL e o CLF-1 apresentaram efeitos prÃ-cicatrizante, bem como uma possÃvel aÃÃo anti-inflamatÃria e investigaÃÃes adicionais sÃo necessÃrias para elucidar o mecanismo de aÃÃo no processo de cicatrizaÃÃo cutÃnea.CoordenaÃÃo de AperfeÃoamento de Pessoal de NÃvel Superior http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=15556application/pdfinfo:eu-repo/semantics/openAccessporreponame:Biblioteca Digital de Teses e Dissertações da UFCinstname:Universidade Federal do Cearáinstacron:UFC2019-01-21T11:28:46Zmail@mail.com -
dc.title.en.fl_str_mv Healing activity assessement of the ethanolic extract and Triterpene 3&#914;, 6 &#914;, 16 &#914; Trihydroxylup-20 (29) â Eno from combretum leprosum leafs in the treatment of cutaneos lesions induced in mice
dc.title.alternative.pt.fl_str_mv AvaliaÃÃo da atividade cicatrizante do extrato etanÃlico e do Triterpeno 3, 6 , 16 Trihydroxylup-20 (29) - Eno provenientes de folhas de Combretum leprosum mart. no tratamento de lesÃes cutÃneas induzidas em camundongos
title Healing activity assessement of the ethanolic extract and Triterpene 3&#914;, 6 &#914;, 16 &#914; Trihydroxylup-20 (29) â Eno from combretum leprosum leafs in the treatment of cutaneos lesions induced in mice
spellingShingle Healing activity assessement of the ethanolic extract and Triterpene 3&#914;, 6 &#914;, 16 &#914; Trihydroxylup-20 (29) â Eno from combretum leprosum leafs in the treatment of cutaneos lesions induced in mice
LÃvia Coelho de Assis
ANATOMIA PATOLOGICA E PATOLOGIA CLINICA
title_short Healing activity assessement of the ethanolic extract and Triterpene 3&#914;, 6 &#914;, 16 &#914; Trihydroxylup-20 (29) â Eno from combretum leprosum leafs in the treatment of cutaneos lesions induced in mice
title_full Healing activity assessement of the ethanolic extract and Triterpene 3&#914;, 6 &#914;, 16 &#914; Trihydroxylup-20 (29) â Eno from combretum leprosum leafs in the treatment of cutaneos lesions induced in mice
title_fullStr Healing activity assessement of the ethanolic extract and Triterpene 3&#914;, 6 &#914;, 16 &#914; Trihydroxylup-20 (29) â Eno from combretum leprosum leafs in the treatment of cutaneos lesions induced in mice
title_full_unstemmed Healing activity assessement of the ethanolic extract and Triterpene 3&#914;, 6 &#914;, 16 &#914; Trihydroxylup-20 (29) â Eno from combretum leprosum leafs in the treatment of cutaneos lesions induced in mice
title_sort Healing activity assessement of the ethanolic extract and Triterpene 3&#914;, 6 &#914;, 16 &#914; Trihydroxylup-20 (29) â Eno from combretum leprosum leafs in the treatment of cutaneos lesions induced in mice
author LÃvia Coelho de Assis
author_facet LÃvia Coelho de Assis
author_role author
dc.contributor.advisor1.fl_str_mv Edson Holanda Teixeira
dc.contributor.advisor1ID.fl_str_mv 71755985304
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/8800103074322967
dc.contributor.referee1.fl_str_mv Max Victor Carioca Freitas
dc.contributor.referee1ID.fl_str_mv 67953662353
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/2609171032311880
dc.contributor.referee2.fl_str_mv Maria Jania Teixeira
dc.contributor.referee2ID.fl_str_mv 23305908300
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.jsp?id=K4798130Z1
dc.contributor.referee3.fl_str_mv Helcio Silva dos Santos
dc.contributor.referee3ID.fl_str_mv 77045050330
dc.contributor.referee3Lattes.fl_str_mv http://lattes.cnpq.br/3478842054984497
dc.contributor.authorID.fl_str_mv 02898042307
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/7104770080836632
dc.contributor.author.fl_str_mv LÃvia Coelho de Assis
contributor_str_mv Edson Holanda Teixeira
Max Victor Carioca Freitas
Maria Jania Teixeira
Helcio Silva dos Santos
dc.subject.cnpq.fl_str_mv ANATOMIA PATOLOGICA E PATOLOGIA CLINICA
topic ANATOMIA PATOLOGICA E PATOLOGIA CLINICA
dc.description.sponsorship.fl_txt_mv CoordenaÃÃo de AperfeÃoamento de Pessoal de NÃvel Superior
dc.description.abstract.por.fl_txt_mv Wound healing is a complex and dynamic process involving an integrated action of several cell types, extracellular matrix and soluble mediators such as cytokines, which serve to recover the integrity of damaged anatomical structure. Natural products are the basis for many biotechnological inputs, as they are biologically active compounds with great potential for therapeutics and can provide molecular basis for many drugs in the clinics. Combretum leprosum Mart. is a native species of Caatinga, important biome in the Brazilian semiarid region, being popularly used in Brazil for the treatment of many diseases such as skin diseases. This study aimed to investigate the pro-healing potential of the ethanol extract (EECL) and the bioactive triterpene 3&#946;, 6 &#946;, 16 &#946;-trihydroxylup-20 (29) ene (CLF-1) isolated from leaves of C. leprosum. Cell migration assay was performed using fibroblasts, which were treated with CLF-1 (1.75 Âg / ml and 2.5 Âg / ml) and analyzed after 0, 6 and 24 hours. Skin wounds (1 cm2) were induced in the dorsal region of mice (n = 30) and were treated with daily topical application (100Âl) according to the group: saline, EECL (100&#956;g / ml) and CLF-1 (100&#956;g / ml) for 12 days. The results showed that CLF-1 did not show toxicity to fibroblasts and further increased cell proliferation and migration so that a significant increase on injury closing percentage was observed using both concentrations (1.75 mg / ml vs 2.5 mg / mL,) compared to control (p <0.05). Within 24 hours, there was also a significant increase in the rate of migration comparing CLF-1 at 2.5 ug / ml and control group (p<0.05). For the in vivo analysis, on the 2nd post surgical day (PS), the lesions treated with EECL and CLF-1 showed moderate presence of vessels in granulation tissue progressing to the dermis, as well as a slight inflammatory infiltrate. It was also observed that, on the 7th PS day, the animals treated with CLF-1 showed moderate angiogenesis, whereas animals treated with saline showed cell proliferation and migration into the wound. Furthermore, on the 12th day PS, the animals treated with EECL and CLF-1 showed complete closing of the injury, while those receiving saline exhibited open wounds. Thus, it was concluded that EECL and CLF-1 showed pro-healing effects, as well as a possible anti-inflammatory action and additional research is needed to elucidate its mechanisms in the wound healing process.
A cicatrizaÃÃo cutÃnea à um processo complexo e dinÃmico que envolve uma aÃÃo integrada de vÃrios tipos celulares, matriz extracelular e mediadores solÃveis como citocinas, que tem por finalidade a recuperaÃÃo da integridade da estrutura anatÃmica danificada. Produtos naturais sÃo a base para diversos insumos biotecnolÃgicos, pois sÃo compostos biologicamente ativos com grande potencial terapÃutico, e que podem formar bases moleculares para muitas drogas em uso clÃnico. Combretum leprosum Mart. à uma espÃcie nativa da Caatinga, bioma importante na regiÃo do semiÃrido brasileiro, sendo esta espÃcie popularmente utilizada no Brasil para o tratamento de diversas doenÃas, tais como doenÃas de pele. O presente estudo investigou o potencial prÃ-cicatrizante do extrato etanÃlico (EECL) e do triterpeno bioativo 3&#946;, 6 &#946;, 16 &#946;-trihydroxylup-20 (29)-eno (CLF-1) isolado das folhas de C. leprosum. Para isto, o ensaio de migraÃÃo de cÃlulas foi realizado com fibroblastos, que foram pÃs tratados com CLF-1 (1,75 &#956;g/mL e 2,5 &#956;g/mL) e analisados em 0, 6 e 24 horas. Feridas cutÃneas (1 cm2) foram realizadas na zona dorsal de camundongos (n=30) e em seguida foram tratadas com uma aplicaÃÃo tÃpica diÃria (100 &#956;l) de acordo com o grupo: salina, EECL (100&#956;g/mL) e CLF-1 (100&#956;g/mL) durante 12 dias. Os resultados demonstraram que CLF-1 nÃo apresentou toxicidade aos fibroblastos e, ainda, em 6 horas estimulou a proliferaÃÃo e migraÃÃo celular, de modo que observou-se um aumento significativo na porcentagem de fechamento em ambas concentraÃÃes (1,75 &#956;g/mL e 2,5 &#956;g/mL) em relaÃÃo ao controle (p<0,05). Em 24 horas, verificou-se tambÃm aumento significativo na taxa fechamento quando comparado CLF-1 2,5 &#956;g/mL em relaÃÃo ao controle (p<0,05). Na avaliaÃÃo in vivo, observou-se no 2 dia pÃs cirÃrgico (P.C.), que as lesÃes tratadas com EECL e CLF-1 mostraram presenÃa moderada de vasos no tecido de granulaÃÃo, progredindo na derme, bem como apresentaram um leve infiltrado inflamatÃrio. Observou-se, ainda, que no 7 dia P.C. os animais tratados com CLF-1 demonstraram moderada angiogÃnese, enquanto que os animais tratados com salina apresentaram proliferaÃÃo e migraÃÃo celular em direÃÃo ao leito da ferida. Ademais, no 12 dia P.C., os animais tratados com EECL e CLF-1 apresentaram o fechamento completo da lesÃo, enquanto os que receberam salina, apresentaram, ainda, feridas abertas. Desta forma, foi possÃvel concluir que o EECL e o CLF-1 apresentaram efeitos prÃ-cicatrizante, bem como uma possÃvel aÃÃo anti-inflamatÃria e investigaÃÃes adicionais sÃo necessÃrias para elucidar o mecanismo de aÃÃo no processo de cicatrizaÃÃo cutÃnea.
description Wound healing is a complex and dynamic process involving an integrated action of several cell types, extracellular matrix and soluble mediators such as cytokines, which serve to recover the integrity of damaged anatomical structure. Natural products are the basis for many biotechnological inputs, as they are biologically active compounds with great potential for therapeutics and can provide molecular basis for many drugs in the clinics. Combretum leprosum Mart. is a native species of Caatinga, important biome in the Brazilian semiarid region, being popularly used in Brazil for the treatment of many diseases such as skin diseases. This study aimed to investigate the pro-healing potential of the ethanol extract (EECL) and the bioactive triterpene 3&#946;, 6 &#946;, 16 &#946;-trihydroxylup-20 (29) ene (CLF-1) isolated from leaves of C. leprosum. Cell migration assay was performed using fibroblasts, which were treated with CLF-1 (1.75 Âg / ml and 2.5 Âg / ml) and analyzed after 0, 6 and 24 hours. Skin wounds (1 cm2) were induced in the dorsal region of mice (n = 30) and were treated with daily topical application (100Âl) according to the group: saline, EECL (100&#956;g / ml) and CLF-1 (100&#956;g / ml) for 12 days. The results showed that CLF-1 did not show toxicity to fibroblasts and further increased cell proliferation and migration so that a significant increase on injury closing percentage was observed using both concentrations (1.75 mg / ml vs 2.5 mg / mL,) compared to control (p <0.05). Within 24 hours, there was also a significant increase in the rate of migration comparing CLF-1 at 2.5 ug / ml and control group (p<0.05). For the in vivo analysis, on the 2nd post surgical day (PS), the lesions treated with EECL and CLF-1 showed moderate presence of vessels in granulation tissue progressing to the dermis, as well as a slight inflammatory infiltrate. It was also observed that, on the 7th PS day, the animals treated with CLF-1 showed moderate angiogenesis, whereas animals treated with saline showed cell proliferation and migration into the wound. Furthermore, on the 12th day PS, the animals treated with EECL and CLF-1 showed complete closing of the injury, while those receiving saline exhibited open wounds. Thus, it was concluded that EECL and CLF-1 showed pro-healing effects, as well as a possible anti-inflammatory action and additional research is needed to elucidate its mechanisms in the wound healing process.
publishDate 2015
dc.date.issued.fl_str_mv 2015-08-28
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publisher.none.fl_str_mv Universidade Federal do CearÃ
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