Author:
Passos Maria Juliane,Chaves Hellíada V.,Barbosa Francisco G.,Mafezoli Jair,Silva-Filho Carlos J. A.,Capistrano André Luiz de O.,Freire Jordânia M. O.,de Souza Nayara A.,Júnior Manoel V. N.,dos Reis Tiago S.,Costa José Jackson do N.,Pereira Karuza Maria A.,de Souza Tamiris de Fátima G.,Rabelo Liviane Maria A.,de Alencar Nylane Maria N.,de Moraes Maria Elisabete A.,Braz Helyson Lucas B.,Goes Paula,Bezerra Mirna Marques
Abstract
Abstract
Background and aim: Combretum leprosum Mart. serves as a medicinal plant in traditional Brazilian medicine. The beneficial effects of C. leprosum Mart. are attributed to the triterpene, 3β,6β,16β-trihydroxylup-20(29)-ene (CL-1). Herein we evaluate the toxicity of two semi-synthetic derivatives from CL-1 (CL-P2 and CL-P2A) in vitro and in vivo, and determine the efficacy in zymosan-induced writhing response and the putative mechanism of action.
Experimental procedure: Toxicity prediction was assessed using the PROTOX-II and ADMETlab 2.0 prediction tools, and SMILES codes for structure identification. In vitro cytotoxicity of the derivatives was tested using the sulforhodamine B assay in L929 and HaCaT cells at 24, 48, and 72 h. Mice received (oral gavage) CL-P2 or CL-P2A (10 mg/kg/d) for 14 days in in vivo toxicity assays. Blood samples and organs (stomach, liver, and kidneys) were collected for AST/ALT level determination and H&E staining, respectively. The anti-nociceptive effect of CL-P2 and CL-P2A (0.1, 1, or 10 mg/kg) was evaluated in the zymosan-induced writhing response. The peritoneal exudate was collected to determine myeloperoxidase (MPO) and superoxide dismutase (SOD) activity, and nitrite concentration.
Results: CL-P-2 and CL-P2A derivatives exhibited low cytotoxicity and did not change body mass, AST/ALT levels, or organ weight. The histopathologic analysis did not reveal significant changes in organs. Both derivatives inhibited the writhing response in a dose-dependent manner. In addition, both derivatives failed to reduce MPO activity. However, CL-P2A increased SOD activity and CL-P2 decreased nitrite/nitrate levels.
Conclusion: CL-P2 and CL-P2A were shown to exhibit anti-nociceptive effects without toxicity. Our data suggest that CL-P2 and CL-P2A efficacy is mediated, at least in part, via antioxidant activity by modulating nitrite/nitrate levels and SOD activity, respectively.