Investigação do potencial antiepiléptico de um óleo de extrato de Cannabis sativa rico em canabidiol: uma abordagem in sílico e in vivo
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFPB |
Texto Completo: | https://repositorio.ufpb.br/jspui/handle/123456789/30081 |
Resumo: | Epilepsy is one of the most common brain diseases, affecting about 2% of the Brazilian population and approximately 50 million people worldwide. Although there is already a considerable number of antiepileptic drugs on the market, there is a need for new drugs, mainly to reduce refractoriness to treatments. In the search for new pharmacological therapies are the substances found in Cannabis sativa. Thus, this work aimed to evaluate the effect of Cannabis sativa rich in CBD (OERCBD) in silico and in vivo. Previously, phytochemical analysis was performed to identify the primary components of OERCBD, followed by molecular docking to evaluate the interaction of these molecules and their ligands. The in vivo studies used male Swiss albino mice (Mus musculus), around 3 months old, which were submitted to behavioral pharmacological screening protocols and determination of LD50 (n=3), induction of seizures by intraperitoneal injection (i.p) of pentylenetetrazole (PTZ, 75mg/kg) (n=8), and the maximum atrial electroshock (MES) model (n=8) with acute or repeated doses administration of OERCBD for 14 days. The animals were divided into a control group, which received olive oil (AZT), groups treated with OERCBD at doses of 15, 30 and 60mg/kg (v.o) and diazepam (DZP, 4mg/kg i.p) in the PTZ test, or phenytoin (FEN, 30mg/kg i.p) in the MES. For the statistical analysis of the induced epileptic seizure tests, the Shapiro-Wilk and Bartlett tests were performed to determine that all parameters should be analyzed using non-parametric tests, followed by the Kruskal-Wallis test and the Dunn test. For the statistical analysis of the percentage of mortality/survival, Fisher's exact test was used, p values <0.05 were considered statistically significant. As a result, gas chromatography-mass spectrometry (GC-MS) identified CBD as the primary component of OERCBD. The molecular docking study demonstrated a good interaction between CBD and the GABAA receptor, but even better with NaV channels. In acute toxicity and LD50, the dose of 300mg/kg (v.o) showed signs of irritability and sedation, while in the doses defined for the treatment (15, 30 and 60mg/kg) such effects were not observed. In the PTZ test, OERCBD in the three doses was not able to increase the latency time for myoclonic and tonic-clonic epileptic seizures, prevent or decrease the severity of the seizures, but protected the animals from the occurrence of death when compared to the group AZT. In MES, the three doses of OERCBD both in acute and repeated doses administration were not able to reduce the duration of tonic epileptic seizures with the extension of the limbs, nor the latency for posture recovery, but in acute administration, there was a reduction in the occurrence of deaths, especially in the highest doses. In the repeated doses administration, there was a worsening in this parameter, mainly in the lowest dose. The in vivo results demonstrate that although there is a good interaction between CBD and GABAA receptors and NaV sodium channels, this may not be enough, reflecting the need for coadministration with other antiepileptic drugs, as already recommended in the clinic. However, the results contribute to understanding the complex brain response to phytocannabinoids and highlight the importance of further investigations to determine your potential pharmacological responses. |
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Investigação do potencial antiepiléptico de um óleo de extrato de Cannabis sativa rico em canabidiol: uma abordagem in sílico e in vivoEpilepsiaCannabis sativaCanabidiolDocking molecularEpilepsyMolecular dockingCNPQ::CIENCIAS HUMANAS::PSICOLOGIAEpilepsy is one of the most common brain diseases, affecting about 2% of the Brazilian population and approximately 50 million people worldwide. Although there is already a considerable number of antiepileptic drugs on the market, there is a need for new drugs, mainly to reduce refractoriness to treatments. In the search for new pharmacological therapies are the substances found in Cannabis sativa. Thus, this work aimed to evaluate the effect of Cannabis sativa rich in CBD (OERCBD) in silico and in vivo. Previously, phytochemical analysis was performed to identify the primary components of OERCBD, followed by molecular docking to evaluate the interaction of these molecules and their ligands. The in vivo studies used male Swiss albino mice (Mus musculus), around 3 months old, which were submitted to behavioral pharmacological screening protocols and determination of LD50 (n=3), induction of seizures by intraperitoneal injection (i.p) of pentylenetetrazole (PTZ, 75mg/kg) (n=8), and the maximum atrial electroshock (MES) model (n=8) with acute or repeated doses administration of OERCBD for 14 days. The animals were divided into a control group, which received olive oil (AZT), groups treated with OERCBD at doses of 15, 30 and 60mg/kg (v.o) and diazepam (DZP, 4mg/kg i.p) in the PTZ test, or phenytoin (FEN, 30mg/kg i.p) in the MES. For the statistical analysis of the induced epileptic seizure tests, the Shapiro-Wilk and Bartlett tests were performed to determine that all parameters should be analyzed using non-parametric tests, followed by the Kruskal-Wallis test and the Dunn test. For the statistical analysis of the percentage of mortality/survival, Fisher's exact test was used, p values <0.05 were considered statistically significant. As a result, gas chromatography-mass spectrometry (GC-MS) identified CBD as the primary component of OERCBD. The molecular docking study demonstrated a good interaction between CBD and the GABAA receptor, but even better with NaV channels. In acute toxicity and LD50, the dose of 300mg/kg (v.o) showed signs of irritability and sedation, while in the doses defined for the treatment (15, 30 and 60mg/kg) such effects were not observed. In the PTZ test, OERCBD in the three doses was not able to increase the latency time for myoclonic and tonic-clonic epileptic seizures, prevent or decrease the severity of the seizures, but protected the animals from the occurrence of death when compared to the group AZT. In MES, the three doses of OERCBD both in acute and repeated doses administration were not able to reduce the duration of tonic epileptic seizures with the extension of the limbs, nor the latency for posture recovery, but in acute administration, there was a reduction in the occurrence of deaths, especially in the highest doses. In the repeated doses administration, there was a worsening in this parameter, mainly in the lowest dose. The in vivo results demonstrate that although there is a good interaction between CBD and GABAA receptors and NaV sodium channels, this may not be enough, reflecting the need for coadministration with other antiepileptic drugs, as already recommended in the clinic. However, the results contribute to understanding the complex brain response to phytocannabinoids and highlight the importance of further investigations to determine your potential pharmacological responses.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESA epilepsia é uma das doenças cerebrais mais comuns, chegando a afetar cerca 2% da população brasileira e aproximadamente 50 milhões de pessoas em todo o mundo. Apesar de já existir um número considerável de fármacos antiepilépticos no mercado, há uma necessidade de novos medicamentos, principalmente para diminuição da refratariedade aos tratamentos. Na busca por novos tratamentos farmacológicos estão as substâncias encontradas na Cannabis sativa. Assim, este trabalho teve como objetivo avaliar o efeito do óleo de extrato da Cannabis sativa rico em CBD (OERCBD) in silico e in vivo. Previamente foi realizada a análise fitoquímica para identificação dos componentes majoritários do OERCBD, seguido do docking molecular para avaliação da interação dessas moléculas e seus ligantes. Os estudos in vivo utilizaram camundongos albinos Swiss (Mus musculus) machos, com cerca de 3 meses de idade, que foram submetidos a avaliação de toxicidade aguda e aos protocolos de triagem farmacológica comportamental (n=3), indução de convulsões pela injeção intraperitoneal (i.p.) de pentilenotetrazol (PTZ, 75mg/kg) (n=8) e Eletrochoque Auricular Máximo (MES) (n=8) com administração aguda ou em repetidas doses do OERCBD durante 14 dias. Os animais foram divididos em grupo controle, que recebeu azeite (AZT), grupos tratados com OERCBD nas doses de 15, 30 e 60mg/kg (v.o.) e diazepam (DZP, 4mg/kg i.p.) no teste do PTZ, ou fenitoína (FEN, 30mg/kg i.p.) no MES. Para análise estatística dos testes de crises epilépticas induzidas, foram realizados os testes de Shapiro-Wilk e de Bartlett para determinar que todos os parâmetros deveriam ser analisados por meio de testes não paramétricos, seguidos do teste de Kruskal-Wallis e teste de Dunn. Para a análise estatística da porcentagem de mortalidade/sobrevivência foi utilizado o teste exato de Fisher, em que valores de p<0,05 foram considerados estatisticamente significativos. Como resultado, a Cromatografia Gasosa Acoplada à Espectrometria de Massas (CG-EM) identificou o CBD como componente majoritário do OERCBD. O estudo docking molecular demonstrou haver boa interação entre o CBD e o receptor GABAA, mas ainda melhor com os canais NaV. Na avaliação da toxicidade aguda, a dose de 300mg/kg (v.o.) apresentou sinais de irritabilidade e sedação, enquanto nas doses definidas para o tratamento (15, 30 e 60mg/kg) tais efeitos não foram observados. No teste do PTZ, o OERCBD nas três doses não foi capaz de aumentar o tempo da latência para as crises epilépticas mioclônicas e tônico-clônicas, impedir ou diminuir a gravidade das crises, mas protegeu os animais da ocorrência de morte quando comparado ao grupo AZT. No MES, as três doses de OERCBD tanto na administração aguda quanto em doses repetidas não foram capazes de diminuir a duração das crises epilépticas tônicas com a extensão dos membros, tampouco a latência para recuperação da postura, porém na administração aguda houve uma redução da ocorrência de mortes, em especial nas doses mais altas, enquanto na administração em doses repetidas houve uma piora em tal parâmetro, principalmente na dose mais baixa. Os resultados in vivo demonstram que apesar de haver uma boa interação do CBD aos receptores GABAA e canais de sódio NaV isso pode não ser suficiente, refletindo a necessidade de coadministração com outros fármacos antiepilépticos. Os resultados obtidos contribuem para a compreensão da complexa resposta cerebral aos fitocanabinoides e destaca a importância de investigações adicionais para determinar suas potenciais as respostas farmacológicas.Universidade Federal da ParaíbaBrasilPsicologiaPrograma de Pós-Graduação em Neurociência Cognitiva e ComportamentoUFPBSalvadori, Mirian Graciela Da Silva Stiebbehttp://lattes.cnpq.br/2669989944106416Santos, Aline Matilde Ferreira dos2024-04-23T10:10:09Z2023-09-132024-04-23T10:10:09Z2023-06-19info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttps://repositorio.ufpb.br/jspui/handle/123456789/30081porAttribution-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nd/3.0/br/info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFPBinstname:Universidade Federal da Paraíba (UFPB)instacron:UFPB2024-04-24T06:10:17Zoai:repositorio.ufpb.br:123456789/30081Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2024-04-24T06:10:17Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false |
dc.title.none.fl_str_mv |
Investigação do potencial antiepiléptico de um óleo de extrato de Cannabis sativa rico em canabidiol: uma abordagem in sílico e in vivo |
title |
Investigação do potencial antiepiléptico de um óleo de extrato de Cannabis sativa rico em canabidiol: uma abordagem in sílico e in vivo |
spellingShingle |
Investigação do potencial antiepiléptico de um óleo de extrato de Cannabis sativa rico em canabidiol: uma abordagem in sílico e in vivo Santos, Aline Matilde Ferreira dos Epilepsia Cannabis sativa Canabidiol Docking molecular Epilepsy Molecular docking CNPQ::CIENCIAS HUMANAS::PSICOLOGIA |
title_short |
Investigação do potencial antiepiléptico de um óleo de extrato de Cannabis sativa rico em canabidiol: uma abordagem in sílico e in vivo |
title_full |
Investigação do potencial antiepiléptico de um óleo de extrato de Cannabis sativa rico em canabidiol: uma abordagem in sílico e in vivo |
title_fullStr |
Investigação do potencial antiepiléptico de um óleo de extrato de Cannabis sativa rico em canabidiol: uma abordagem in sílico e in vivo |
title_full_unstemmed |
Investigação do potencial antiepiléptico de um óleo de extrato de Cannabis sativa rico em canabidiol: uma abordagem in sílico e in vivo |
title_sort |
Investigação do potencial antiepiléptico de um óleo de extrato de Cannabis sativa rico em canabidiol: uma abordagem in sílico e in vivo |
author |
Santos, Aline Matilde Ferreira dos |
author_facet |
Santos, Aline Matilde Ferreira dos |
author_role |
author |
dc.contributor.none.fl_str_mv |
Salvadori, Mirian Graciela Da Silva Stiebbe http://lattes.cnpq.br/2669989944106416 |
dc.contributor.author.fl_str_mv |
Santos, Aline Matilde Ferreira dos |
dc.subject.por.fl_str_mv |
Epilepsia Cannabis sativa Canabidiol Docking molecular Epilepsy Molecular docking CNPQ::CIENCIAS HUMANAS::PSICOLOGIA |
topic |
Epilepsia Cannabis sativa Canabidiol Docking molecular Epilepsy Molecular docking CNPQ::CIENCIAS HUMANAS::PSICOLOGIA |
description |
Epilepsy is one of the most common brain diseases, affecting about 2% of the Brazilian population and approximately 50 million people worldwide. Although there is already a considerable number of antiepileptic drugs on the market, there is a need for new drugs, mainly to reduce refractoriness to treatments. In the search for new pharmacological therapies are the substances found in Cannabis sativa. Thus, this work aimed to evaluate the effect of Cannabis sativa rich in CBD (OERCBD) in silico and in vivo. Previously, phytochemical analysis was performed to identify the primary components of OERCBD, followed by molecular docking to evaluate the interaction of these molecules and their ligands. The in vivo studies used male Swiss albino mice (Mus musculus), around 3 months old, which were submitted to behavioral pharmacological screening protocols and determination of LD50 (n=3), induction of seizures by intraperitoneal injection (i.p) of pentylenetetrazole (PTZ, 75mg/kg) (n=8), and the maximum atrial electroshock (MES) model (n=8) with acute or repeated doses administration of OERCBD for 14 days. The animals were divided into a control group, which received olive oil (AZT), groups treated with OERCBD at doses of 15, 30 and 60mg/kg (v.o) and diazepam (DZP, 4mg/kg i.p) in the PTZ test, or phenytoin (FEN, 30mg/kg i.p) in the MES. For the statistical analysis of the induced epileptic seizure tests, the Shapiro-Wilk and Bartlett tests were performed to determine that all parameters should be analyzed using non-parametric tests, followed by the Kruskal-Wallis test and the Dunn test. For the statistical analysis of the percentage of mortality/survival, Fisher's exact test was used, p values <0.05 were considered statistically significant. As a result, gas chromatography-mass spectrometry (GC-MS) identified CBD as the primary component of OERCBD. The molecular docking study demonstrated a good interaction between CBD and the GABAA receptor, but even better with NaV channels. In acute toxicity and LD50, the dose of 300mg/kg (v.o) showed signs of irritability and sedation, while in the doses defined for the treatment (15, 30 and 60mg/kg) such effects were not observed. In the PTZ test, OERCBD in the three doses was not able to increase the latency time for myoclonic and tonic-clonic epileptic seizures, prevent or decrease the severity of the seizures, but protected the animals from the occurrence of death when compared to the group AZT. In MES, the three doses of OERCBD both in acute and repeated doses administration were not able to reduce the duration of tonic epileptic seizures with the extension of the limbs, nor the latency for posture recovery, but in acute administration, there was a reduction in the occurrence of deaths, especially in the highest doses. In the repeated doses administration, there was a worsening in this parameter, mainly in the lowest dose. The in vivo results demonstrate that although there is a good interaction between CBD and GABAA receptors and NaV sodium channels, this may not be enough, reflecting the need for coadministration with other antiepileptic drugs, as already recommended in the clinic. However, the results contribute to understanding the complex brain response to phytocannabinoids and highlight the importance of further investigations to determine your potential pharmacological responses. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-09-13 2023-06-19 2024-04-23T10:10:09Z 2024-04-23T10:10:09Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufpb.br/jspui/handle/123456789/30081 |
url |
https://repositorio.ufpb.br/jspui/handle/123456789/30081 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nd/3.0/br/ info:eu-repo/semantics/openAccess |
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Attribution-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nd/3.0/br/ |
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openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal da Paraíba Brasil Psicologia Programa de Pós-Graduação em Neurociência Cognitiva e Comportamento UFPB |
publisher.none.fl_str_mv |
Universidade Federal da Paraíba Brasil Psicologia Programa de Pós-Graduação em Neurociência Cognitiva e Comportamento UFPB |
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reponame:Biblioteca Digital de Teses e Dissertações da UFPB instname:Universidade Federal da Paraíba (UFPB) instacron:UFPB |
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Universidade Federal da Paraíba (UFPB) |
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UFPB |
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UFPB |
reponame_str |
Biblioteca Digital de Teses e Dissertações da UFPB |
collection |
Biblioteca Digital de Teses e Dissertações da UFPB |
repository.name.fl_str_mv |
Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB) |
repository.mail.fl_str_mv |
diretoria@ufpb.br|| diretoria@ufpb.br |
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1801843027465994240 |