Oxidative stress-mediated bimodal regulation of polymorphonuclear leukocyte spreading by polyphenolic compounds.

Pyrogallol-bearing polyphenolic compounds induce spreading of polymorphonuclear leukocytes (PMNL), although their optimal concentrations for induction of spreading are quite different (2000, 200, and 2 µM for pyrogallol, (-)-epigallocatechin gallate (EGCG), and tannic acid (TA), respectively), and TA tends to inhibit spreading at higher concentrations. In this study, we examined the involvement of oxidative stress in the regulation of PMNL spreading by these compounds. All three compounds in solution generated H(2)O(2) to a similar extent. Adsorption of the polyphenols to cell surfaces and their accumulation within cells were assessed by detection of the H(2)O(2) precursor O(2)(-) produced by the compounds through reduction of cytochrome c and p-nitro-blue tetrazolium, respectively. TA showed the highest degree of adsorption. EGCG adhered only to PMNL pre-fixed by paraformaldehyde, whereas pyrogallol did not adhere. None of the compounds caused intracellular O(2)(-) generation. A non-pyrogallic compound, 1,2,4-benzenetriol (BT), also produced H(2)O(2); it had no stimulatory effect on PMNL spreading, but inhibited spreading induced by other stimuli. BT did not adhere to PMNL but accumulated within them, and generated O(2)(-) in the presence of glycine. Thiol antioxidants abrogated all of the above spreading-regulatory effects of the polyphenolic compounds. We conclude that H(2)O(2)-generating polyphenols bimodally regulate the spreading of PMNL by subjecting them to oxidative stress. The ability of polyphenol to adhere to, or accumulate within, PMNL may govern the nature of the oxidative stress and determine the optimal concentration of each compound for induction of spreading, as well as whether spreading is promoted or inhibited.

Biphasic regulation of polymorphonuclear leukocyte spreading by polyphenolic compounds with pyrogallol moieties.

Green tea polyphenols have been reported to have anti-inflammatory activities, although the molecular mechanisms responsible for this effect remain unclear. In the present study, we examined the effect of green tea extract and a variety of polyphenolic compounds on spreading of peripheral blood polymorphonuclear leukocytes (PMNs) over fibrinogen-coated surfaces. Green tea extract exerted a biphasic effect on PMN spreading; it induced or suppressed spreading at low and high concentrations, respectively. We also found that pyrogallol-bearing compounds have spreading induction activity. Among the compounds tested, tannic acid (TA) had the strongest activity; the concentrations required for induction of maximal spreading were 2 microM for TA, 200 microM for (-)-epigallocatechin gallate, and 2000 microM for the other active compounds. Furthermore, TA was the only compound showing a biphasic effect similar to that of green tea extract; TA at 20 or 200 microM suppressed spreading. The spreading-stimulatory signal was still latent during PMN exposure to TA at concentrations that inhibited spreading, because the pre-exposed PMNs underwent spreading when plated after removal of free TA by centrifugation. The spreading-inhibitory effect of TA at high concentrations overcame the induction of spreading by other stimuli, including phorbol 12-myristate 13-acetate, hydrogen peroxide, denatured fibrinogen surfaces, and naked plastic surfaces. These results suggest that TA as well as green tea extract is bi-functional, having pro-inflammatory and anti-inflammatory effects at low and high concentrations, respectively. Pharmacological use of TA may thus provide new strategies aimed at regulation of PMN spreading for control of inflammation.