Evaluation of Antiradical and Anti-Inflammatory Activities of Ethyl Acetate and Butanolic Subfractions of Agelanthus dodoneifolius (DC.) Polhill & Wiens (Loranthaceae) Using Equine Myeloperoxidase and Both PMA-Activated Neutrophils and HL-60 Cells.

Rainatou Boly,Jacques Dubois,Innocent Pierre Guissou,Ange Mouithys-Mickalad,Marius Lompo,Thierry Franck,Stephan Kohnen,Didier Serteyn

doi:10.1155/2015/707524

Abstract

The ethyl acetate and n-butanolic subfractions of Agelanthus dodoneifolius were investigated for their antioxidant and antimyeloperoxidase (MPO) activities. The reactive oxygen species (ROS) generation was assessed by lucigenin-enhanced chemiluminescence (CL) and dichlorofluorescein- (DCF-) induced fluorescence techniques from phorbol myristate acetate- (PMA-) stimulated equine neutrophils and human myeloid cell line HL-60, respectively. In parallel, the effects of the tested subfractions were evaluated on the total MPO release by stimulated neutrophils and on the specific MPO activity by means of immunological assays. The results showed the potent activity of the butanolic subfraction, at least in respect of the chemiluminescence test (IC50 = 0.3 ± 0.1 µg/mL) and the ELISA and SIEFED assays (IC50 = 2.8 ± 1.2 µg/mL and 1.3 ± 1.0 µg/mL), respectively. However, the ethyl acetate subfraction was found to be the most potent in the DCF assay as at the highest concentration, DCF fluorescence intensity decreases of about 50%. Moreover, we demonstrated that the ethyl acetate subfraction was rich in catechin (16.51%) while it was not easy to identify the main compounds in the butanolic subfraction using the UPLC-MS/MS technique. Nevertheless, taken together, our results provide evidence that Agelanthus dodoneifolius subfractions may represent potential sources of natural antioxidants and of antimyeloperoxidase compounds.

Highlights

Free radicals, especially reactive oxygen species (ROS), which are produced during the oxidative burst of polymorphonuclear neutrophils (PMNs), are generated by two key enzymes, namely, NADPH oxidase and myeloperoxidase (MPO) [1]
The reactive oxygen species (ROS) generation was assessed by lucigenin-enhanced chemiluminescence (CL) and dichlorofluorescein- (DCF-) induced fluorescence techniques from phorbol myristate acetate- (PMA-) stimulated equine neutrophils and human myeloid cell line HL-60, respectively
Our work is in this context of standardization through purification of Agelanthus dodoneifolius (Loranthaceae) extracts, a plant used in Burkina Faso folk medicine for the treatment of cardiovascular and respiratory diseases, stomachache, and wounds

Summary

Introduction

Especially reactive oxygen species (ROS), which are produced during the oxidative burst of polymorphonuclear neutrophils (PMNs), are generated by two key enzymes, namely, NADPH oxidase and myeloperoxidase (MPO) [1]. PMNs are thought to produce both extracellular and intracellular reactive oxygen species that may have an important role in the host defense against invading microorganisms [2]. Myeloperoxidase (MPO), a heme-containing peroxidase, is released outside cellular compartment and is tightly linked to both inflammation and oxidative stress. In conditions of excessive and uncontrolled inflammation, the degranulation as well as release of MPO in the extracellular space has been linked to the pathogenesis of various diseases such as atherosclerosis, multiple sclerosis, and cancer [5]. Compounds with radical scavenging and antimyeloperoxidase activities may be relevant in the treatment of diseases where free radicals and MPO are implicated

Methods

Results

Conclusion