Evaluation of The Effects of Oxidative Stress, Lipid Peroxidation, Anti-hyperglycemic, Cardiovascular and Anti-Pulmonary Inflammation Markers on In Vivo Experimental Models of the Natural Diterpene trans-Dehydrocrotonin Unload and Encapsulated into SNEDDS Colloidal System
Croton cajucara Benth; trans-Dehydrocrotonin; Copaifera reticulata Ducke; SNEDDS system; Oxidative stress; Hypoglycemic activity; Pulmonary anti-inflammatory activity.
Croton cajucara Benth, widely used in folk medicine in the Amazon region of Brazil, is widely used to treat diabetes, diarrhea, fever, jaundice, hepatitis and malaria, among other pharmaceutical indications. In this work, the diterpene 19-nor-clerodane trans-dehydrocrotonin (t-DCTN), isolated from this Croton, was co-encapsulated with Copaiferareticulata Ducke oil-resin (OCPR) in a self-nanoemulsifying drug delivery system (SNEDDS) aiming of evaluating its efficacy in an experimental model of type 1 diabetes mellitus (DM1) and a model of pulmonary inflammation by lipopolysaccharide (LPS) of Escherichia coli O55:B5, on biochemical parameters, oxidative stress and lipid peroxidation. Rattus norvegicus Wistar animals were kept under temperature conditions (24 ± 1ºC), light/dark cycle (12h), with free access to feed. After fasting (12 h), they received 45 mg kg-1 (i.p.) of STZ diluted in citrate buffer pH 4.5. For the lung inflammation (PI) model, female Wistar rats (200 g to 300 g) that received LPS instillation (4 mg kg-1) (i.t.) were used. The tested samples were: solution of the unload t-DCTN solubilized in DMSO (15 mg mL-1), the carrier system containing OCPR (0.5%) so-called SNEDDS-OCPR, and the nanoproduct resulting from the co-encapsulation of the oil OCPR and the diterpene t-DCTN (SNEDDS-OCPR-DCTN). The diabetic animals were treated (v.o.) with SNEDDS-OCPR-DCTN at the doses 0.1 mL 100 g-1 and 0.05 mL 100 g-1, as well the unload t-DCTN at the unique dose 0,1 mL 100 g-1, and SNEDDS-OCPR at the unique dose 0.1 mL 100 g-1. At the end of treatment (28 days), hematological and biochemical parameters, markers of oxidative stress and lipid peroxidation were determined, in addition to histopathological analysis. In the DM1A model, the hypoglycemic activity of the nanoproduct SNEDDS-OCPR-DCTN was evidenced only at the dose 0.1 mL 100 g-1, with antioxidant potential due to the reduction of oxidative stress and lipid peroxidation. In this model, in the groups treated with SNEDDS-OCPR-DCTN and the unload t-DCTN. For all treated animal groups was evidenced variation in the weight, increased water and feed intake, as well as reduction in glycemic indexes. Increases in AST, PGT, and plasma urea were observed in all treatment groups, but the reduction in LDH was observed for SNEDDS-OCPR-DCTN group; neutrophilia was observed in the group treated with the unload t-DCTN, and lymphopenia in the group treated with SNEDDS-OCPR-DCTN. Reduction in oxidative stress (MDA and NO) was observed in the group treated with the unload t-DCTN. For all treated groups, histopathological alterations were observed in the liver tissue, but in the renal tissue, alterations were observed in the groups treated with SNEDDS-OCPR (moderate for both nephrosis and passive congestion) and SNEDDS-OCPR-DCTN (moderate for both mixed inflammatory infiltrate and passive congestion). For the PI model, no variation in the weight of the animals was observed, and a lower leukocyte index was observed in the group treated with the unload t-DCTN. The elevation of cardiac markers was more evident in the group treated with SNEDDS-OCPR-DCTN (0.1 mL 100 g-1) and less evident in the unload t-DCTN group. The group treated with SNEDDS-OCPR-DCTN (0.1 mL 100 g-1) showed reduced oxidative stress in cardiac (MDA) and pulmonary (CAT) tissue. The colloidal nanoproduct SNEDDS-OCPR-DCTN is suitable for oral ingestion, showed effectiveness as hypoglycemic and anti-inflammatory pulmonary, and is herein considered as an advanced colloidal (SNEDDS) system to optimize the availability of the bioactives OCPR and t-DCTN.