Blue biotechnology on the Coast of Portugal: a prospection of neurological bioactive compounds from marine sponges
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10362/164461 |
Resumo: | Marine environments are vastly unexplored territories, being considered an untapped resource of chemical and biological molecules. Chances of new opportunities are emerging and likewise, the dis- covery of new potential pharmaceuticals. These bioactive compounds are present in a variety of inver- tebrates, such as the sponges (Porifera) studied in this project, found on the Coast of Sagres, south of Portugal. The extracts and fractions obtained from this organism were submitted to neuronal bioactivity evaluations by determining their possible effects on both potassium (K+) and sodium (Na+) currents of small diameter dorsal root ganglion neurons (sdDRG), dissected from naïve rats. The most active sam- ple in assessed resorting to electrophysiology studies, was elected to undergo bio guided fractionation process, to reach a purified fraction (99% purity) of the most prominent peak (S4U1). Under the presence of S4U1, whole-cell voltage-clamp were conducted to screen S4U1 against a platform of voltage-gated K+ and Na+ channels present at the surface of a well-established pain sensing cell model: sdDRG neurons (ex vivo testing). Ex vivo experiments recordings in rat sdDRG neurons showed decreased mean current density values in the presence of S4U1, mostly observed in the slow component of K+ currents (IK), suggesting a specific effect over IK, and in peak Na+ current. These results might be chiefly supported by a facilitation of the inactivation of K+ and Na+ currents, observed in the hyperpolarizing shift of both steady state inactivation curves, and in the increasing rate of the decaying phase of K+ currents. Thus, by ensuring the absence of S4U1 toxicity in human and animal cell lines (MTS), the bio- technological value of S4U1 was further analyzed through a study of its potential analgesic effect in animal (chronic) pain models (Chronic Constriction Injury- CCI; in vivo testing). Through the intrave- nous administration of S4U1 (in vivo testing) the reduction of pain was effectively observed, since, following an evaluation method in animal models, %MPE, there was an increase in the force applied by the von Frey monofilaments (vFF) in the ipsilateral side, in each period of time. The results showed a neuronal bioactive effect of S4U1 on pain sensing model cell (sdDRG), which was translated into a potential analgesic value verified in chronic pain models. Despite, the need to increase the experimental sample size, S4U1 is here presented as a possible new therapeutic drug for chronic pain treatment. |
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Blue biotechnology on the Coast of Portugal: a prospection of neurological bioactive compounds from marine spongesSpongesNeuronal BioactivityAnalgesic PotentialElectrophysiologyDomínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e TecnologiasMarine environments are vastly unexplored territories, being considered an untapped resource of chemical and biological molecules. Chances of new opportunities are emerging and likewise, the dis- covery of new potential pharmaceuticals. These bioactive compounds are present in a variety of inver- tebrates, such as the sponges (Porifera) studied in this project, found on the Coast of Sagres, south of Portugal. The extracts and fractions obtained from this organism were submitted to neuronal bioactivity evaluations by determining their possible effects on both potassium (K+) and sodium (Na+) currents of small diameter dorsal root ganglion neurons (sdDRG), dissected from naïve rats. The most active sam- ple in assessed resorting to electrophysiology studies, was elected to undergo bio guided fractionation process, to reach a purified fraction (99% purity) of the most prominent peak (S4U1). Under the presence of S4U1, whole-cell voltage-clamp were conducted to screen S4U1 against a platform of voltage-gated K+ and Na+ channels present at the surface of a well-established pain sensing cell model: sdDRG neurons (ex vivo testing). Ex vivo experiments recordings in rat sdDRG neurons showed decreased mean current density values in the presence of S4U1, mostly observed in the slow component of K+ currents (IK), suggesting a specific effect over IK, and in peak Na+ current. These results might be chiefly supported by a facilitation of the inactivation of K+ and Na+ currents, observed in the hyperpolarizing shift of both steady state inactivation curves, and in the increasing rate of the decaying phase of K+ currents. Thus, by ensuring the absence of S4U1 toxicity in human and animal cell lines (MTS), the bio- technological value of S4U1 was further analyzed through a study of its potential analgesic effect in animal (chronic) pain models (Chronic Constriction Injury- CCI; in vivo testing). Through the intrave- nous administration of S4U1 (in vivo testing) the reduction of pain was effectively observed, since, following an evaluation method in animal models, %MPE, there was an increase in the force applied by the von Frey monofilaments (vFF) in the ipsilateral side, in each period of time. The results showed a neuronal bioactive effect of S4U1 on pain sensing model cell (sdDRG), which was translated into a potential analgesic value verified in chronic pain models. Despite, the need to increase the experimental sample size, S4U1 is here presented as a possible new therapeutic drug for chronic pain treatment.Os ambientes marinhos apesar de serem considerados como recursos naturais de grande relevân- cia para moléculas químicas e biológicas, são ainda territórios vastamente inexplorados. Contudo, no- vas oportunidades estão a surgir relativamente ao interesse marítimo e, consequentemente, à descoberta de potenciais novos produtos farmacêuticos. Estes compostos bioativos estão presentes numa variedade de invertebrados, tais como as esponjas (Porifera), podendo ser encontradas ao largo da Costa de Sagres, sul de Portugal e, intrinsecamente ligadas ao propósito desta dissertação. Os extratos e frações obtidos de um organismo colhido foram submetidos a avaliações da bioatividade neuronal para determinar pos- síveis efeitos tanto em correntes de potássio (K+) como de sódio (Na+) em small diameter Dorsal Root Ganlions (sdDRG) neurons, dissecados de ratos naïve. A amostra eleita com a mais proeminente em termos de bioatividade, no decorrer das análises eletrofisiológicas, foi subjugada a um processo de fra- cionamento bioguiado, com a intenção de atingir uma fração purificada (99% de pureza) do pico mais saliente (S4U1). Posteriormente, estudos de whole-cell voltage clamp, recorrendo ao mesmo modelo celular (tes- tes ex vivo), foram realizados com e sem a presença de S4U1, estabelecendo assim uma compreensão mais concreta dos seus efeitos nos canais de K+e Na+. Assim, com registos experimentais foi possível observar um decréscimo de valores médios de densidade de corrente aquando presença de S4U1, ob- servados principalmente na componente lenta das correntes K+ (IK) e nas correntes de Na+, sugerindo tanto um possível efeito específico em IK como no pico das correntes de Na+. Após uma análise deta- lhada dos resultados foi também possível cimentar a sugestão anteriormente mencionada, relativamente à existência de efeitos a nível da neuroexcitabilidade dos canais, complementada pela facilidade obser- vada de inativação das correntes de K+ e Na+, através de uma consequente deslocação das curvas de inativação para estados mais hiperpolarizantes. Tais conclusões permitiram inferir um possível efeito terapêutico em modelos de dor. Deste modo, ao garantir a inexistência de toxicidade de S4U1 em linhas celulares humanas e animais (MTS), o valor biotecnológico de S4U1 foi aprofundado através de um posterior estudo do seu potencial efeito analgésico em modelos de dor (crónica) em animal (Chronic Constriction Injury- CCI; testes in vivo). Através da administração intravenosa de S4U1(testes in vivo) a redução de dor foi efeti- vamente observada, uma vez que, seguindo um método de avaliação em modelos animais, %MPE, registou-se um aumento da força aplicada pelos monofilamentos de von Frey (vFF) no lado ipsilateral, num determinado período de tempo. Os resultados demonstram um potencial bioativo neuronal associado a S4U1 em modelos celu- lares de sensibilidade à dor (sdDRG), que foi traduzido num potencial valor analgésico verificado em modelos de dor crónica. Apesar da necessidade de aumentar o tamanho da amostra experimental, o S4U1 é aqui apresentado como um possível novo medicamento com fins terapêuticos para o tratamento da dor crónica.Lima, PedroBastos, AndréRUNGonçalves, Afonso Miguel dos Santos2024-03-05T17:36:34Z2022-072022-07-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/164461enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:52:31Zoai:run.unl.pt:10362/164461Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T04:00:11.674744Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Blue biotechnology on the Coast of Portugal: a prospection of neurological bioactive compounds from marine sponges |
title |
Blue biotechnology on the Coast of Portugal: a prospection of neurological bioactive compounds from marine sponges |
spellingShingle |
Blue biotechnology on the Coast of Portugal: a prospection of neurological bioactive compounds from marine sponges Gonçalves, Afonso Miguel dos Santos Sponges Neuronal Bioactivity Analgesic Potential Electrophysiology Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
title_short |
Blue biotechnology on the Coast of Portugal: a prospection of neurological bioactive compounds from marine sponges |
title_full |
Blue biotechnology on the Coast of Portugal: a prospection of neurological bioactive compounds from marine sponges |
title_fullStr |
Blue biotechnology on the Coast of Portugal: a prospection of neurological bioactive compounds from marine sponges |
title_full_unstemmed |
Blue biotechnology on the Coast of Portugal: a prospection of neurological bioactive compounds from marine sponges |
title_sort |
Blue biotechnology on the Coast of Portugal: a prospection of neurological bioactive compounds from marine sponges |
author |
Gonçalves, Afonso Miguel dos Santos |
author_facet |
Gonçalves, Afonso Miguel dos Santos |
author_role |
author |
dc.contributor.none.fl_str_mv |
Lima, Pedro Bastos, André RUN |
dc.contributor.author.fl_str_mv |
Gonçalves, Afonso Miguel dos Santos |
dc.subject.por.fl_str_mv |
Sponges Neuronal Bioactivity Analgesic Potential Electrophysiology Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
topic |
Sponges Neuronal Bioactivity Analgesic Potential Electrophysiology Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
description |
Marine environments are vastly unexplored territories, being considered an untapped resource of chemical and biological molecules. Chances of new opportunities are emerging and likewise, the dis- covery of new potential pharmaceuticals. These bioactive compounds are present in a variety of inver- tebrates, such as the sponges (Porifera) studied in this project, found on the Coast of Sagres, south of Portugal. The extracts and fractions obtained from this organism were submitted to neuronal bioactivity evaluations by determining their possible effects on both potassium (K+) and sodium (Na+) currents of small diameter dorsal root ganglion neurons (sdDRG), dissected from naïve rats. The most active sam- ple in assessed resorting to electrophysiology studies, was elected to undergo bio guided fractionation process, to reach a purified fraction (99% purity) of the most prominent peak (S4U1). Under the presence of S4U1, whole-cell voltage-clamp were conducted to screen S4U1 against a platform of voltage-gated K+ and Na+ channels present at the surface of a well-established pain sensing cell model: sdDRG neurons (ex vivo testing). Ex vivo experiments recordings in rat sdDRG neurons showed decreased mean current density values in the presence of S4U1, mostly observed in the slow component of K+ currents (IK), suggesting a specific effect over IK, and in peak Na+ current. These results might be chiefly supported by a facilitation of the inactivation of K+ and Na+ currents, observed in the hyperpolarizing shift of both steady state inactivation curves, and in the increasing rate of the decaying phase of K+ currents. Thus, by ensuring the absence of S4U1 toxicity in human and animal cell lines (MTS), the bio- technological value of S4U1 was further analyzed through a study of its potential analgesic effect in animal (chronic) pain models (Chronic Constriction Injury- CCI; in vivo testing). Through the intrave- nous administration of S4U1 (in vivo testing) the reduction of pain was effectively observed, since, following an evaluation method in animal models, %MPE, there was an increase in the force applied by the von Frey monofilaments (vFF) in the ipsilateral side, in each period of time. The results showed a neuronal bioactive effect of S4U1 on pain sensing model cell (sdDRG), which was translated into a potential analgesic value verified in chronic pain models. Despite, the need to increase the experimental sample size, S4U1 is here presented as a possible new therapeutic drug for chronic pain treatment. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-07 2022-07-01T00:00:00Z 2024-03-05T17:36:34Z |
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 |
http://hdl.handle.net/10362/164461 |
url |
http://hdl.handle.net/10362/164461 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
dc.source.none.fl_str_mv |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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