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.

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.

Pharmacological properties and related constituents of stem bark of Pterocarpus erinaceus Poir. (Fabaceae)

Objective To screen methanol and dichloromethane extracts of stem bark of Pterocarpus erinaceus for anti-inflammatory, analgesic, in vitro antioxidant activities and phytochemical analysis. Methods Anti-inflammatory activity was determined by using carrageenan induced-edema of mice paw and croton oil-induced edema of mice ear; analgesic effect was evaluated using acetic acid-induced writhing. Phytochemical screening of extracts was performed by thin layer chromatography. The chromatographic fractionation led to the isolation of main active components as friedelin, lupeol and epicathechin. The structures were established by TLC and nuclear magnetic resonance studies. Results Both methanol and dichloromethane extracts, friedelin, lupeol and epicatechin showed a significant anti-inflammatory effect using croton oil induced-ear edema. Furthermore, the action of dichloromethane extract was more important. At the doses of 100 and 200 mg/kg, the methanol extract was able to reduce the carrageenan induced-hind paw edema, while at the doses of 100, 200 and 400 mg/kg, it showed an important analgesic effect against writhing induced by acetic acid injection of 38.8%, 68.0% and 74.3%, respectively. Antioxidative properties of methanol extract and its dichloromethane and ethyl acetate fractions were assessed by using the 1,1-diphenyl-2-picrylhydrazyl method. The methanol extract showed the stronger radical scavenging activity than dichloromethane and ethyl acetate fractions, with an antiradical power of 5, 3.5 and 2 respectively. The main components isolated from these extracts as friedelin, lupeol and epicathechin were responsible of these activities. Conclusions The results suggest that the stem bark extracts of Pterocarpus erinaceus possessed important anti-inflammatory, analgesic activities and strong antioxidant properties, therefore, they could be used as natural potential ingredients for pharma ceutical industry.

Pharmacological properties and related constituents of stem bark of Pterocarpus erinaceus Poir. (Fabaceae)

OBJECTIVE@#To screen methanol and dichloromethane extracts of stem bark of Pterocarpus erinaceus for anti-inflammatory, analgesic, in vitro antioxidant activities and phytochemical analysis.@*METHODS@#Anti-inflammatory activity was determined by using carrageenan induced-edema of mice paw and croton oil-induced edema of mice ear; analgesic effect was evaluated using acetic acid-induced writhing. Phytochemical screening of extracts was performed by thin layer chromatography. The chromatographic fractionation led to the isolation of main active components as friedelin, lupeol and epicathechin. The structures were established by TLC and nuclear magnetic resonance studies.@*RESULTS@#Both methanol and dichloromethane extracts, friedelin, lupeol and epicatechin showed a significant anti-inflammatory effect using croton oil induced-ear edema. Furthermore, the action of dichloromethane extract was more important. At the doses of 100 and 200 mg/kg, the methanol extract was able to reduce the carrageenan induced-hind paw edema, while at the doses of 100, 200 and 400 mg/kg, it showed an important analgesic effect against writhing induced by acetic acid injection of 38.8%, 68.0% and 74.3%, respectively. Antioxidative properties of methanol extract and its dichloromethane and ethyl acetate fractions were assessed by using the 1,1-diphenyl-2-picrylhydrazyl method. The methanol extract showed the stronger radical scavenging activity than dichloromethane and ethyl acetate fractions, with an antiradical power of 5, 3.5 and 2 respectively. The main components isolated from these extracts as friedelin, lupeol and epicathechin were responsible of these activities.@*CONCLUSIONS@#The results suggest that the stem bark extracts of Pterocarpus erinaceus possessed important anti-inflammatory, analgesic activities and strong antioxidant properties, therefore, they could be used as potential natural ingredients in the pharmaceutical industry.

Modulatory activities of Agelanthus dodoneifolius (Loranthaceae) extracts on stimulated equine neutrophils and myeloperoxidase activity.

Agelanthus dodoneifolius DC Danser (Loranthaceae) is used for the treatment of various diseases including asthma. The aqueous and hydroalcoholic extracts have been reported to have anti-inflammatory, spasmolytic and bronchorelaxant activities. The present study investigates the effects of the aqueous decoction and the diethyl ether, ethyl acetate and butanolic fractions of Agelanthus dodoneifolius DC Danser (Loranthaceae) on reactive oxygen species (ROS) production and myeloperoxidase (MPO) release by phorbol 12-myristate 13-acetate (PMA)-stimulated equine neutrophils and on purified equine MPO activity. ROS production and MPO release by the PMA-stimulated neutrophils were measured by the lucigenin-enhanced chemiluminescence and ELISA assays, respectively. Specific immunological extraction followed by enzymatic detection (SIEFED) was used to specifically measure the equine MPO activity. Identification and quantification of the individual and total phenolic and flavonoid compounds were performed using UPLC-MS/MS equipment and colorimetric methods involving Folin-Ciocalteu and AlCl3, respectively. All the tested extracts displayed dose-dependent inhibitory effects on the oxidant activities of neutrophils; a stronger effect was observed with the organic fractions than the aqueous decoction. These findings could be correlated with a high content of phenolic and flavonoid compounds. The results confirm the previously shown anti-inflammatory effect of Agelanthus dodoneifolius and its potential use for the treatment of neutrophil-dependent inflammatory diseases.