Apoptose em macr??fagos induzida por nanoc??psulas com n??cleo lip??dico (LNC) carregadas com diclofenaco

• Author(s): Barbosa, Aguyda Rayany Cavalcante
• Date: 2019
• Origin: Oasisbr
• Subject(s): Macr??fagos; Agentes antiinflamat??rios; Artrite reumat??ide; Apoptose; CI??NCIAS BIOL??GICAS: IMUNOLOGIA; Diclofenaco; Nanoc??psula; Macr??fago; Inflama????o
• Document Type: Master thesis
• Language: Portuguese

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Rejected by Divis??o de Documenta????o/BC Biblioteca Central ([e-mail]), reason: O dep??sito necessita das seguintes altera????es: 1. Corrigir Ficha Catalogr??fica. No campo "nomes e primeiros sobrenomes" n??o deve constar o seu ??ltimo sobrenome (Barbosa), pois h?? um campo espec??fico para ele (??ltimo sobrenome). Desta forma a Ficha deve ser "Barbosa, Aguyda Rayany Cavalcante". A Ficha pode ser elaborada em https://biblioteca.ufam.edu.br/servicos/elaboracao-de-ficha-catalografica 2. Corrigir a Carta de Encaminhamento para o Autodep??sito. O nome do orientador foi substitu??do por c??digos. O modelo da Carta de Encaminhamento para o Autodep??sito est?? dispon??vel em: https://biblioteca.ufam.edu.br/servicos/teses-e-dissertacoes on 2019-11-14T19:56:20Z (GMT)

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Made available in DSpace on 2019-11-19T20:57:43Z (GMT). No. of bitstreams: 4 Ata de Defesa Mestrado.jpeg: 123596 bytes, checksum: 79605551f5c05404bb676e261f919aba (MD5) Dissertac??ao.pdf: 9244401 bytes, checksum: ff54ef2860e663f64ad0f8bd9d367b73 (MD5) Ficha catalografica.pdf: 1903 bytes, checksum: 608eb833c8ac684c4c177194f9b83476 (MD5) CartaEncaminhamentoAutodepo??sito-2.pdf: 208160 bytes, checksum: 15afd62fa1ce86e80ef58fd14a985d00 (MD5) Previous issue date: 2016-10-07

FAPEAM - Funda????o de Amparo ?? Pesquisa do Estado do Amazonas

(92) 994595440

Diclofenac is a nonsteroidal anti-inflammatory drug, used to treat inflammatory muscle skeletal diseases, including the autoimmune diseases as Rheumatoid Arthritis (RA). In chronic synovitis of Rheumatoid Arthritis the macrophages play a central role in inflammation and the induction of apoptosis in those cells can represent a central paper in the treatment. For this work was used a formula of Diclofenac carried in nanocapsules of lipid core (LNC-DIC), validating the paper of this formula in apoptosis induction in macrophages. The LNC was formed parting from a organogel core, coated with a polymeric poly (e-caprolactona) wall and stabilized with Polyssorbato 80, obtaining a diameter D [4,3] of 243nm for DIC and 319nm for LNC-DIC. By that way were used the formulas Diclofenac (DIC), LNC-DIC and only LNC in human macrophages (THP-1) and murines (J774). The IC50% for toxicity in macrophages for the formulations was 646??M for DIC, 144,5??M for LNC-DIC in THP-1, 251,7??M for DIC and 24,35??M for LNC-DIC in J774. The formula LNC-DIC reduced strongly the mitochondrial membrane potential related to DIC (-44,65??3,36 e 21,79??2,19, p<0,0001, respectively) in THP-1, besides the J774 (-5,65??0,88 e -26,59??7,21, p<0,0001).There was also a ROS intracellular bigger production when compared with DIC in THP-1 lineage (27,24??0,90 e 36,88??2,49, p=0,0002). The LNC-DIC formulation increased the percentage of cells with activated caspasa 3 when compared with DIC (53,08??6,23 e 81,70??9,09, p=0,0007) and the caspasa 3 activity in the studied concentration. Besides this, LNC-DIC induced higher percentage of Annexin V+ cells compared to DIC (6,26??0,34 e 12,68??0,22, p=0,0001). When was validated the DNA damage index using comet test, there was not any difference between the tested DIC and LNC-DIC (44,24??2,65 e 53,33??4,16, p=0,586). Taking together, those results show that nanoencapsulated formulation LNC-DIC has an apoptosis inductor action, independent of DNA damage. Possibly related to intracellular ROS production and endoplasmic reticule stress, opening the possibilities for a functional and ship treatment for RA, through the use of nanocapsules of lipid core carried with Diclofenac.

O Diclofenaco ?? uma droga anti-inflamat??ria n??o esteroide (AINE), utilizada no tratamento de afec????es inflamat??rias musculoesquel??ticas, incluindo as doen??as autoimunes como a Artrite Reumatoide (AR). Na sinovite cr??nica da AR os macr??fagos exercem papel central na inflama????o e a indu????o de apoptose destas c??lulas pode vir a representar um papel central no tratamento. Neste trabalho, utilizou-se de uma formula????o de diclofenaco carregada em nanoc??psulas de n??cleo lip??dico (LNC-DIC), avaliando-se o papel desta formula????o na indu????o de apoptose em macr??fagos. A LNC foi formada a partir de um n??cleo organogel, revestido com uma parede polim??rica poly(e- caprolactona) e estabilizada com polyssorbato 80, obtendo di??metro D[4,3] de 243nm para DIC e 319nm para LNC-DIC. Dessa forma foram utilizadas as formula????es Diclofenaco (DIC), LNC-DIC e apenas LNC em macr??fagos humanos (THP-1) e Murinos (J774). A IC50% para toxicidade em macr??fagos para as formula????es foi 646??M para DIC e 144,5??M para LNC-DIC em THP-1 e 251,7??M para DIC e 24,35??M para LNC-DIC em J774. A formula????o LNC-DIC reduziu acentuadamente o potencial de membrana mitocondrial em rela????o a DIC (-44,65??3,36 e 21,79??2,19, p<0,0001, respectivamente) em THP-1, al??m de J774 (-5,65??0,88 e -26,59??7,21, p<0,0001). Tamb??m houve maior produ????o de ROS intracelular para LNC-DIC em rela????o a DIC na linhagem THP-1 (27,24??0,90 e 36,88??2,49, p=0,0002). A formula????o LNC-DIC aumentou o percentual de c??lulas com caspase-3 ativa comparada ?? DIC (53,08??6,23 e 81,70??9,09, p=0,0007) e a atividade de caspase-3 nas concentra????es estudadas. Al??m disso, LNC-DIC induziu maior percentual de c??lulas Anexina V+ comparada a DIC (6,26??0,34 e 12,68??0,22, p=0,0001). Quando avaliado o ??ndice de dano ao DNA pelo ensaio cometa, n??o houve diferen??a entre as formula????es testadas DIC e LNC-DIC (44,24??2,65 e 53,33??4,16, p=0,586). Tomados em conjunto, esses resultados demonstram que a formula????o nanoencapsulada LNC-DIC tem a????o indutora de apoptose, de forma independente do dano ao DNA, possivelmente relacionada a produ????o de ROS intracelular e/ou estresse do ret??culo endoplasm??tico, abrindo assim a possibilidade de um tratamento eficaz e barato para a AR, atrav??s do uso de Nanoc??psulas de N??cleo Lip??dico carregado com Diclofenaco.