The natural stilbenoid pinosylvin and activated neutrophils: effects on oxidative burst, protein kinase C, apoptosis and efficiency in adjuvant arthritis

Abstract

Aim: To investigate the effects of the naturally occurring stilbenoid pinosylvin on neutrophil activity in vitro and in experimental arthritis, and to examine whether protein kinase C (PKC) activation served as an assumed target of pinosylvin action. Methods: Fresh human blood neutrophils were isolated. The oxidative burst of neutrophils was evaluated on the basis of enhanced chemiluminescence. Neutrophil viability was evaluated with flow cytometry, and PKC phosphorylation was assessed by Western blotting analysis. Adjuvant arthritis was induced in Lewis rats with heat-killed Mycobacterium butyricum, and the animals were administered with pinosylvin (30 mg/kg, po) daily for 21 d after arthritis induction. Results: In isolated human neutrophils, pinosylvin (10 and 100 µmol/L) significantly decreased the formation of oxidants, both extraand intracellularly, and effectively inhibited PKC activation stimulated by phorbol myristate acetate (0.05 µmol/L). The inhibition was not due to neutrophil damage or increased apoptosis. In arthritic rats, the number of neutrophils in blood was dramatically increased, and whole blood chemiluminescence (spontaneous and PMA-stimulated) was markedly enhanced. Pinosylvin administration decreased the number of neutrophils (from 69 671±5588/µL to 51 293±3947/µL, P=0.0198) and significantly reduced the amount of reactive oxygen species in blood. Conclusion: Pinosylvin is an effective inhibitor of neutrophil activity, and is potentially useful as a complementary medicine in states associated with persistent inflammation.

Water-Soluble Components of Pinus pinaster Wood

“…A wide scope of applications has been suggested or pinosylvins including antifungal and antibacterial agents (Lindberg et al 2004;Lee et al 2005;Celimene et al 1999;Gref et al 2000;Venäläinen et al 2004) with activity towards Listeria monocytogenes (Gözü et al 2010), cytotoxicity against a murine hepatic carcinoma cell line (Välimaa et al 2007), antimetastatic activity (Park et al 2012), antiinflammatory action based on the reduction of blood reactive species (Jančinová et al 2012;Bauerova et al 2011), and angiogenic effects (Kimura and Sumiyoshi 2011).…”

Equol Effectively Inhibits Toxic Activity of Human Neutrophils without Influencing Their Viability

“…Neutrophils were washed with PBS and centrifuged (10 min, 170 g , RT). Neutrophils were adjusted to a final concentration of 10 4 cells/1 μl [23] . The final suspension contained more than 96% of viable cells and was used maximally for 2 h -as long as control chemiluminescence kept constant.…”

“…Western blot analysis was performed as previously described [23,25] . Isolated human neutrophils (2.5 × 10 6 cells) were incubated at 37 ° C with equol (10 or 100 μmol/l, 1 min), stimulated with PMA (0.15 μmol/l, 1 min) and lysed by sonication in a lysing solution containing protease and phosphatase inhibitors.…”

Selective inhibition of extracellular oxidants liberated from human neutrophils—A new mechanism potentially involved in the anti-inflammatory activity of hydroxychloroquine

“…Each experimental group consisted of 10 animals and the substances tested were applied over a period 21 days from arthritis initiation. Then the formation of reactive oxygen species (spontaneous and stimulated with PMA) was determined on the basis of luminolenhanced chemiluminescence [25,26]. The samples contained 50 μl aliquots of 1.25 mmol/l luminol, 40 U/ml horseradish peroxidase, rat blood diluted 200-times with Tyrode's solution, 0.05 μmol/l PMA (or PBS) and Tyrode's solution.…”