Atividade larvicida e inibição in vitro de proteases intestinais de Aedes aegypti por extratos de sementes de plantas da caatinga

Arboviruses are epidemiologically relevant viruses transmitted by arthropods, affecting especially tropical countries, such as Brazil. Currently, the fight against arboviruses is carried out mainly through population control of Aedes aegypti, main urban vector mosquito. Recently, the search for eco...

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Autor principal: Bezerra, Pedro Vítor Vale
Outros Autores: Uchôa, Adriana Ferreira
Formato: bachelorThesis
Idioma:pt_BR
Publicado em: Universidade Federal do Rio Grande do Norte
Assuntos:
Homogenato intestinal
Zimograma
Atividade aedicida
Proteases serínicas
Intestinal homogenate
Zymography
Aedicidal activity
Serine proteases
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIA
Endereço do item:https://repositorio.ufrn.br/handle/123456789/49312
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id ri-123456789-49312
record_format dspace
institution Repositório Institucional
collection RI - UFRN
language pt_BR
topic Homogenato intestinal
Zimograma
Atividade aedicida
Proteases serínicas
Intestinal homogenate
Zymography
Aedicidal activity
Serine proteases
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIA
spellingShingle Homogenato intestinal
Zimograma
Atividade aedicida
Proteases serínicas
Intestinal homogenate
Zymography
Aedicidal activity
Serine proteases
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIA
Bezerra, Pedro Vítor Vale
Atividade larvicida e inibição in vitro de proteases intestinais de Aedes aegypti por extratos de sementes de plantas da caatinga
description Arboviruses are epidemiologically relevant viruses transmitted by arthropods, affecting especially tropical countries, such as Brazil. Currently, the fight against arboviruses is carried out mainly through population control of Aedes aegypti, main urban vector mosquito. Recently, the search for eco friendly alternatives for vector control has been encouraged, such as the development of botanical insecticides. In this scenario, the Caatinga, a center of biodiversity, represents an important source of plant species for prospecting insecticidal molecules. The present research investigated the potential activity of seed extracts from 3 plant species from the Caatinga, Amburana cearensis, Anadenanthera colubrina and Erythrina velutina, against A. aegypti larvae, and sought to evaluate, in vitro, the ability to inhibit intestinal proteases of the mosquito larvae. Thus, bioassays were performed exposing the larvae to different extract concentrations, and the percentage of larval survival at 24 and 48 hours was analyzed in order to estimate the lethal concentrations 50% (LC50). For the in vitro tests, homogenates were produced from the larvae midguts, in order to analyze the inhibition by exposure to extracts quantitatively, through an enzyme activity assay, and qualitatively, through zymography. The aqueous extracts of A. cearensis (CE), A. colubrina (AE) and E. velutina (ME) compromised larval survival, presenting LC50 equal to 0.67 mg/mL, 0.28 mg/mL and 1, 85 mg/mL, after 24h of exposure, and 0.24 mg/mL, 0.05 mg/mL and 0.91 mg/mL, after 48h of exposure, respectively. The initial characterization of the extracts suggests an efficiency in the extraction conditions, with dry weight yield between 14.53 mg and 34.50 mg, total yield between 7.56% and 54.16%, and soluble protein contents between 1,16 mg/ml and 2,30 mg/ml. The extracts soluble proteins possibly contributed for the enzyme activity impairment of A. aegypti larvae intestinal homogenate proteases, with specific inhibitory activity in CE (662.65 IU/mg), AE (226.19 IU/mg) and ME (496.11 IU/mg). The zymography analysis revealed the presence of proteolytic bands of high relative molecular weight (above 66 kDa), medium (between 45 kDa and 30 kDa) and low (below 30 kDa), and that the extracts affected distinct proteases groups in the larval homogenate. The results indicate the inhibition of intestinal proteases may comprise one of the modes of action for the larvicidal activity of the extracts, especially for the CE extract. Complementary tests are underway to confirm this relationship.
author2 Uchôa, Adriana Ferreira
author_facet Uchôa, Adriana Ferreira
Bezerra, Pedro Vítor Vale
format bachelorThesis
author Bezerra, Pedro Vítor Vale
author_sort Bezerra, Pedro Vítor Vale
title Atividade larvicida e inibição in vitro de proteases intestinais de Aedes aegypti por extratos de sementes de plantas da caatinga
title_short Atividade larvicida e inibição in vitro de proteases intestinais de Aedes aegypti por extratos de sementes de plantas da caatinga
title_full Atividade larvicida e inibição in vitro de proteases intestinais de Aedes aegypti por extratos de sementes de plantas da caatinga
title_fullStr Atividade larvicida e inibição in vitro de proteases intestinais de Aedes aegypti por extratos de sementes de plantas da caatinga
title_full_unstemmed Atividade larvicida e inibição in vitro de proteases intestinais de Aedes aegypti por extratos de sementes de plantas da caatinga
title_sort atividade larvicida e inibição in vitro de proteases intestinais de aedes aegypti por extratos de sementes de plantas da caatinga
publisher Universidade Federal do Rio Grande do Norte
publishDate 2022
url https://repositorio.ufrn.br/handle/123456789/49312
work_keys_str_mv AT bezerrapedrovitorvale atividadelarvicidaeinibicaoinvitrodeproteasesintestinaisdeaedesaegyptiporextratosdesementesdeplantasdacaatinga
AT bezerrapedrovitorvale larvicidalactivityandinvitroinhibitionofaedesaegyptiintestinalproteasesbyseedextractsfromcaatingaplants
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spelling ri-123456789-493122023-01-03T17:12:12Z Atividade larvicida e inibição in vitro de proteases intestinais de Aedes aegypti por extratos de sementes de plantas da caatinga Larvicidal activity and in vitro inhibition of Aedes aegypti intestinal proteases by seed extracts from caatinga plants Bezerra, Pedro Vítor Vale Uchôa, Adriana Ferreira https://orcid.org/0000-0002-8674-6959 http://lattes.cnpq.br/0567529774568955 https://orcid.org/0000-0002-3822-6502 http://lattes.cnpq.br/6644671747055211 Sá, Giulian César da Silva https://orcid.org/0000-0002-4734-4228 http://lattes.cnpq.br/2134578884742038 Santos, Elizeu Antunes dos https://orcid.org/0000-0003-4372-2161 http://lattes.cnpq.br/6762251930590306 Inácio, Cássio Lázaro Silva https://orcid.org/0000-0002-1666-6646 http://lattes.cnpq.br/4910533645616566 Homogenato intestinal Zimograma Atividade aedicida Proteases serínicas Intestinal homogenate Zymography Aedicidal activity Serine proteases CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIA Arboviruses are epidemiologically relevant viruses transmitted by arthropods, affecting especially tropical countries, such as Brazil. Currently, the fight against arboviruses is carried out mainly through population control of Aedes aegypti, main urban vector mosquito. Recently, the search for eco friendly alternatives for vector control has been encouraged, such as the development of botanical insecticides. In this scenario, the Caatinga, a center of biodiversity, represents an important source of plant species for prospecting insecticidal molecules. The present research investigated the potential activity of seed extracts from 3 plant species from the Caatinga, Amburana cearensis, Anadenanthera colubrina and Erythrina velutina, against A. aegypti larvae, and sought to evaluate, in vitro, the ability to inhibit intestinal proteases of the mosquito larvae. Thus, bioassays were performed exposing the larvae to different extract concentrations, and the percentage of larval survival at 24 and 48 hours was analyzed in order to estimate the lethal concentrations 50% (LC50). For the in vitro tests, homogenates were produced from the larvae midguts, in order to analyze the inhibition by exposure to extracts quantitatively, through an enzyme activity assay, and qualitatively, through zymography. The aqueous extracts of A. cearensis (CE), A. colubrina (AE) and E. velutina (ME) compromised larval survival, presenting LC50 equal to 0.67 mg/mL, 0.28 mg/mL and 1, 85 mg/mL, after 24h of exposure, and 0.24 mg/mL, 0.05 mg/mL and 0.91 mg/mL, after 48h of exposure, respectively. The initial characterization of the extracts suggests an efficiency in the extraction conditions, with dry weight yield between 14.53 mg and 34.50 mg, total yield between 7.56% and 54.16%, and soluble protein contents between 1,16 mg/ml and 2,30 mg/ml. The extracts soluble proteins possibly contributed for the enzyme activity impairment of A. aegypti larvae intestinal homogenate proteases, with specific inhibitory activity in CE (662.65 IU/mg), AE (226.19 IU/mg) and ME (496.11 IU/mg). The zymography analysis revealed the presence of proteolytic bands of high relative molecular weight (above 66 kDa), medium (between 45 kDa and 30 kDa) and low (below 30 kDa), and that the extracts affected distinct proteases groups in the larval homogenate. The results indicate the inhibition of intestinal proteases may comprise one of the modes of action for the larvicidal activity of the extracts, especially for the CE extract. Complementary tests are underway to confirm this relationship. CNPq Arboviroses são doenças epidemiologicamente relevantes transmitidas por artrópodes, afetando sobretudo países tropicais, como o Brasil. Atualmente o enfrentamento das arboviroses é realizado, principalmente, através do controle populacional do Aedes aegypti, principal mosquito vetor urbano. Recentemente, a busca por alternativas ecologicamente amigáveis para o controle vetorial tem sido incentivada, como o desenvolvimento de inseticidas botânicos. Nesse cenário, a Caatinga, centro de biodiversidade, representa uma fonte importante de espécies vegetais para prospecção de moléculas inseticidas. A presente pesquisa investigou a potencial atividade de extratos de sementes de 3 espécies vegetais da Caatinga, Amburana cearensis, Anadenanthera colubrina e Erythrina velutina, contra larvas de A. aegypti, e buscou avaliar, in vitro, suas capacidades de inibição das proteases intestinais das larvas deste mosquito. Bioensaios foram realizados expondo as larvas em diferentes concentrações dos extratos, sendo analisada a taxa de sobrevivência larval em 24h e 48h, para estimar as concentrações letais 50% (CL50). Homogenatos foram produzidos a partir dos intestinos médios das larvas para analisar in vitro a inibição pela exposição aos extratos, quantitativamente, através de ensaio de atividade enzimática, e qualitativamente, através de zimogramas. Os extratos aquosos de A. cearensis (EC), A. colubrina (EA) e E. velutina (EM) comprometeram a sobrevivência das larvas, apresentando CL50 iguais a 0,67 mg/mL, 0,28 mg/mL e 1,85 mg/mL, após 24h de exposição, e 0,24 mg/mL, 0,05 mg/mL e 0,91 mg/mL, após 48h de exposição, respectivamente. A caracterização inicial dos extratos sugere uma eficiência nas condições de extrações, com rendimento em peso seco entre 14,53 mg e 34,50 mg, rendimento total entre 7,56% e 54,16%, e teores de proteínas solúveis entre 1,16 mg/mL e 2,30 mg/mL. As proteínas solúveis nos extratos possivelmente favoreceram o comprometimento da atividade enzimática das proteases do homogenato intestinal das larvas de A. aegypti, com atividade inibitória específica em EC (662,65 UI/mg), EA (226,19 UI/mg) e EM (496,11 UI/mg). As análises dos zimogramas revelaram a presença de bandas proteolíticas de peso molecular relativo alto (acima de 66 kDa), médio (entre 45 kDa e 30 Kda) e baixo (abaixo de 30 kDa), e que os extratos afetaram grupos distintos de proteases do homogenato das larvas. Os resultados indicam que a inibição das proteases intestinais pode compreender um dos modos de ação para a atividade larvicida dos extratos, em especial para o extrato EC. Testes complementares estão em andamento para confirmar essa relação. 2022-09-08T14:27:19Z 2022-09-08T14:27:19Z 2022-07-22 bachelorThesis BEZERRA, Pedro Vítor Vale. Atividade larvicida e inibição in vitro de proteases intestinais de Aedes aegypti por extratos de sementes de plantas da caatinga. 2022. 48 f. Monografia (Graduação em Ciências Biológicas) – Universidade Federal do Rio Grande do Norte, Natal, 2022. https://repositorio.ufrn.br/handle/123456789/49312 pt_BR application/pdf Universidade Federal do Rio Grande do Norte Brasil UFRN Ciências Biológicas Departamento de Biologia Celular e Genética

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