Prostasomes inhibit the NADPH oxidase activity of human neutrophils.
Prostasomes are particular lipid vesicles secreted by the prostate in human semen and involved in several physiological functions such as the improvement of sperm motility or immunomodulation. We have previously shown that they reduced the overall reactive oxygen species (ROS) production of seminal polymorphonuclear neutrophils (PMN). The present study was conducted to define the mechanism by which prostasomes inhibit the ROS production of blood and seminal PMN. The luminol chemiluminescence measuring total ROS production of blood PMN stimulated by either a phorbol ester (PMA) or a chemoattractant peptide, formyl-Met-Leu-Phe (fMLP) was significantly inhibited by prostasomes. The NADPH oxidase activity of the PMN was measured by 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1, 2-a]pyrazin-3-one (MCLA) chemiluminescence. Prostasomes inhibited the NADPH oxidase activity of blood or seminal PMN and increased the lag-phase of the enzyme after PMA stimulation. Prostasomes also inhibited significantly the NADPH oxidase activity of fMLP stimulated blood PMN, but the inhibition was not significant for seminal PMN. The lipid composition of blood PMN was analysed and compared to the lipid composition of prostasomes. This showed that prostasomes had a high cholesterol:phospholipid molar ratio and a high proportion of sphingomyelin. Together with the fact that prostasomes can rigidify the plasma membrane of blood PMN, these results led us to postulate that prostasomes inhibit the NADPH oxidase activity of PMN by lipid transfer from the prostasomes to the plasma membrane of the PMN.
Saez
F
F
Laboratoire de Biologie de la Reproduction - CECOS, and Laboratoire de Biochimie, CHU Hôtel Dieu, 63003 Clermont-Ferrand, France.
Motta
C
C
Boucher
D
D
Grizard
G
G
D002784:Cholesterol / Q000378:metabolism; D005456:Fluorescent Dyes; D006801:Humans; D007093:Imidazoles; D008163:Luminescent Measurements; D008297:Male; D008560:Membrane Fluidity; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors* / Q000097:blood / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology*; D010743:Phospholipids / Q000097:blood / Q000378:metabolism; D011467:Prostate / Q000737:chemistry / Q000378:metabolism*; D011719:Pyrazines; D017382:Reactive Oxygen Species / Q000378:metabolism*; D022142:Secretory Vesicles / Q000378:metabolism*; D012661:Semen / Q000737:chemistry*; D013094:Spermatozoa / Q000378:metabolism; D013696:Temperature; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D005456:Fluorescent Dyes; D007093:Imidazoles; D010743:Phospholipids; D011719:Pyrazines; D017382:Reactive Oxygen Species; C063412:2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo(1,2-alpha)pyrazin-3-one; D009240:N-Formylmethionine Leucyl-Phenylalanine; D002784:Cholesterol; D019255:NADPH Oxidases; D013755:Tetradecanoylphorbol Acetate