Ação do extrato metanólico de Plantago major (tanchagem/tansagem) em micro-organismos de interesse médico
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Data
2022-01-24
Autores
Sousa, Débora Silva Marques de
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Universidade Estadual Paulista (Unesp)
Resumo
A resistência aos fármacos antimicrobianos é um sério problema de saúde pública. Micro-organismos como Staphylococcus aureus resistente à meticilina (MRSA), Pseudomonas aeruginosa, Klebsiella pneumoniae produtora de carbapenemase, Pythium insidiosum e Candida auris, apresentam altas taxas de resistência aos antimicrobianos utilizados para tratar as infecções, por esse motivo, se faz necessário o desenvolvimento de novos fármacos. Os produtos naturais, inclusive as plantas medicinais, têm sido amplamente estudados para o desenvolvimento de fármacos para tratamento de diversas enfermidades. Plantago major é uma planta com diversas propriedades biológicas, incluindo atividade antibacteriana, antifúngica e ação cicatrizante. Assim sendo, nosso objetivo foi estudar o efeito inibidor do extrato metanólico preparado com as folhas desta planta em linhagens de S. aureus resistente à meticilina, S. aureus sensível à meticilina, P. aeruginosa, K. pneumoniae produtora de carbapenemase, C. auris, P. insidiosum, Sporotrhix globosa, Sporotrhrix brasiliensis e Sporotrhirx schenkii. Foram realizados ensaios de microdiluição em placas de ELISA para determinação da Concentração Inibitória Mínima (CIM), Concentração Bactericida (CBM) e Fungicida Mínima (CFM), bem como a verificação de sinergismo do extrato com os antimicrobianos comumente utilizados, através das metodologias de Time kill curve utilizando contagem de colônias e valores de densidade óptica (OD600). Também foi analisada a redução da produção de biofilme por MRSA, P. aeruginosa, K. pneumoniae produtora de carbapenemase e C. auris. Para determinação da CIM e CFM de P., fragmentos padronizados com 5 mm de diâmetro das hifas foram deixados em contato com diferentes concentrações do extrato e posteriormente transferidos para placas de Ágar Sabouraud para avaliação do crescimento por 7 dias. As CIMs variaram entre 2,5 e 45 mg/mL. Foi possível verificar interação sinérgica bactericida do extrato com imipenem frente à K. pneumoniae produtora de carbapenemase, bacteriostático nas combinações do extrato com cefalotina e oxacilina para MRSA e fungistático entre extrato e anfotericina B para C. auris. Foi possível observar, ainda, uma redução de 85% da produção de biofilme por K. pneumoniae produtora de carbapenemase, 90% de redução para MRSA ATCC, 54% para MRSA isolado clínico, 13,7% para P. aeruginosa isolado clínico, 69,3% para C. auris ATCC e 90,78% para C. auris isolado clínico. O extrato metanólico das folhas de Plantago major apresentou ação importante contra os micro-organismos analisados no presente estudo, revelando ser um produto natural potencialmente aplicável à produção de fármacos para utilização terapêutica nas doenças provocadas pelos micro-organismos testados neste estudo.
Antimicrobial drug resistance is a serious public health problem. Microorganisms such as methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, carbapenemase-producing Klebsiella pneumoniae, Pythium insidiosum and Candida auris, have high rates of resistance to antimicrobials used to treat infections, for this reason, it is necessary to develop new drugs. Natural products, including medicinal plants, have been widely studied for the development of drugs for the treatment of several diseases. Plantago major is a plant with several biological properties, including antibacterial, antifungal and healing action. Therefore, our objective was to study the inhibitory effect of the methanolic extract prepared with the leaves of this plant on strains of methicillin-resistant S. aureus, methicillin-sensitive S. aureus, P. aeruginosa, carbapenemase-producing K. pneumoniae, C. auris, P. insidiosum, Sporotrhix globosa, Sporotrhrix brasiliensis and Sporotrhirx schenkii. Microdilution assays were carried out on ELISA plates to determine the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal (MBC) and Fungicide Concentration (MFC), as well as the verification of synergism of the extract with commonly used antimicrobials, through the methodologies of Time kill curve by counting colonies and optical density (OD600). The reduction of biofilm production by MRSA, P. aeruginosa, carbapenemase-producing K. pneumoniae and C. auris was also analyzed. To determine the MIC and MFC for P. insidiosum, standardized fragments with 5 mm of diameter of hyphae were kept in contact with different concentrations of the extract and later transferred to Sabouraud Agar plates for growth evaluation for 7 days. MICs ranged between 2.5 and 45 mg/mL. It was possible to verify synergistic bactericidal interaction of extract with imipenem against carbapenemase-producing K. pneumoniae, bacteriostatic in combinations of extract with cephalotin and oxacillin for MRSA, and fungistatic between extract and amphotericin B for C. auris. It was also possible to observe an 85% reduction in biofilm production by carbapenemase-producing K. pneumoniae, 90% reduction for MRSA ATCC, 54% for isolated MRSA clinical isolate, 13.7% for P. aeruginosa clinical isolated, 69.3% for C. auris ATCC and 90.78% for C. auris clinical isolated. The methanol extract of Plantago major leaves showed important action against the microorganisms analyzed in this study, revealing that it is a natural product potentially applicable to the production of drugs for therapeutic use in the treatment of diseases caused by the microorganisms tested in this study.
Antimicrobial drug resistance is a serious public health problem. Microorganisms such as methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, carbapenemase-producing Klebsiella pneumoniae, Pythium insidiosum and Candida auris, have high rates of resistance to antimicrobials used to treat infections, for this reason, it is necessary to develop new drugs. Natural products, including medicinal plants, have been widely studied for the development of drugs for the treatment of several diseases. Plantago major is a plant with several biological properties, including antibacterial, antifungal and healing action. Therefore, our objective was to study the inhibitory effect of the methanolic extract prepared with the leaves of this plant on strains of methicillin-resistant S. aureus, methicillin-sensitive S. aureus, P. aeruginosa, carbapenemase-producing K. pneumoniae, C. auris, P. insidiosum, Sporotrhix globosa, Sporotrhrix brasiliensis and Sporotrhirx schenkii. Microdilution assays were carried out on ELISA plates to determine the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal (MBC) and Fungicide Concentration (MFC), as well as the verification of synergism of the extract with commonly used antimicrobials, through the methodologies of Time kill curve by counting colonies and optical density (OD600). The reduction of biofilm production by MRSA, P. aeruginosa, carbapenemase-producing K. pneumoniae and C. auris was also analyzed. To determine the MIC and MFC for P. insidiosum, standardized fragments with 5 mm of diameter of hyphae were kept in contact with different concentrations of the extract and later transferred to Sabouraud Agar plates for growth evaluation for 7 days. MICs ranged between 2.5 and 45 mg/mL. It was possible to verify synergistic bactericidal interaction of extract with imipenem against carbapenemase-producing K. pneumoniae, bacteriostatic in combinations of extract with cephalotin and oxacillin for MRSA, and fungistatic between extract and amphotericin B for C. auris. It was also possible to observe an 85% reduction in biofilm production by carbapenemase-producing K. pneumoniae, 90% reduction for MRSA ATCC, 54% for isolated MRSA clinical isolate, 13.7% for P. aeruginosa clinical isolated, 69.3% for C. auris ATCC and 90.78% for C. auris clinical isolated. The methanol extract of Plantago major leaves showed important action against the microorganisms analyzed in this study, revealing that it is a natural product potentially applicable to the production of drugs for therapeutic use in the treatment of diseases caused by the microorganisms tested in this study.
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Palavras-chave
Fitoterapia, Plantago major, Microbiologia médica, Testes de sensibilidade microbiana