Apoptose em macrófagos induzida por nanocápsulas com núcleo lipídico (LNC) carregadas com diclofenaco

Abstract

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 646μM for DIC, 144,5μM for LNC-DIC in THP-1, 251,7μM for DIC and 24,35μM 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.