Effect of Anadenanthera colubrina protease inhibitors as an antiinflamatory mediator

• Published in: Natural product research 2021-05, Vol.35 (10), p.1690-1695
• Main Authors: Guarneire, Gracimério J, Cardoso Junior, Oswaldo, Marques Lima, Nerilson, Aguilar Santos, Emanuel, Aguiar Schulze, Caio, Pereira Silva, Wellington, Pedro Oliveira Batista, João, de Paula Carli, Gabriela, Castro, Sandra Br, Alves, Caio Cesar S, Carli, Alessandra P
• Format: Article
• Language: English
• Subjects: Anadenanthera colubrine; anti-inflammatory; Bioassays; Biocompatibility; Cell viability; Cytotoxicity; Dexamethasone; Enzymatic activity; Gel electrophoresis; Hemagglutination; High-performance liquid chromatography; Inflammation; Liquid chromatography; Protease; Protease inhibitors; Proteinase inhibitors; Proteins; Sodium lauryl sulfate; Toxicity; Trypsin; Tumor necrosis factor-α; Zeta potential
• ISSN: 1478-6419; 1478-6427
• DOI: 10.1080/14786419.2019.1624962

This work aimed to obtain and characterize protease inhibitors from A. colubrina leaves and evaluate their potential as inflammatory mediator and cell viability. The protein extract was analyzed and characterized by SDS-PAGE, RP-HPLC-PDA, MALDI-TOF/MS and Zeta potential. Bioassays were conducted in order to evaluate the cell viability in RAW 264.7, in vitro (NO and TNF-α production inhibition) and in vivo anti-inflammatory potential, inhibition rate of trypsin and hemagglutination activity from protein extract. The results revealed the presence of bands at 14, 21 and 30 kDa in SDS-PAGE, the RP-HPLC-PDA analysis showed peaks at 12, 13, 28 and 40 minutes and MALDI-TOF/MS showed peaks with 3.4, 4.7, 5.6, 9.4 and 11.2 kDa. The protein extracts presented enzymatic activity inhibition of trypsin (IC 50 59.2 μgmL −1 ), did not show any cytotoxicity to RAW264.7 cells, hemagglutination 8HU and insignificant reduction in NO and TNF-α production and reduced anti-inflammatory potential in vivo compared to dexamethasone.