title: The Impact of Myeloperoxidase and Activated Macrophages on Metaphase II Mouse Oocyte Quality.
abstract: Myeloperoxidase (MPO), an abundant heme-containing enzyme present in neutrophils, monocytes, and macrophages, is produced in high levels during inflammation, and associated with poor reproductive outcomes. MPO is known to generate hypochlorous acid (HOCl), a damaging reactive oxygen species (ROS) utilizing hydrogen peroxide (H2O2) and chloride (Cl-). Here we investigate the effect of activated immune cells and MPO on oocyte quality. Mouse metaphase II oocytes were divided into the following groups: 1) Incubation with a catalytic amount of MPO (40 nM) for different incubation periods in the presence of 100 mM Cl- with and without H2O2 and with and without melatonin (100 μM), at 37°C (n = 648/648 total number of oocytes in each group for oocytes with and without cumulus cells); 2) Co-cultured with activated mouse peritoneal macrophage and neutrophils cells (1.0 x 106 cells/ml) in the absence and presence of melatonin (200 μM), an MPO inhibitor/ROS scavenger, for different incubation periods in HTF media, at 37°C (n = 200/200); 3) Untreated oocytes incubated for 4 hrs as controls (n = 73/64). Oocytes were then fixed, stained and scored based on the microtubule morphology and chromosomal alignment. All treatments were found to negatively affect oocyte quality in a time dependent fashion as compared to controls. In all cases the presence of cumulus cells offered no protection; however significant protection was offered by melatonin. Similar results were obtained with oocytes treated with neutrophils. This work provides a direct link between MPO and decreased oocyte quality. Therefore, strategies to decrease MPO mediated inflammation may influence reproductive outcomes.
authors.affiliation: Department of Obstetrics and Gynecology, the C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, MI 48201, United States of America.
authors.forename: Faten
authors.initials: F
authors.lastname: Shaeib
authors.affiliation: Department of Obstetrics and Gynecology, the C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, MI 48201, United States of America.
authors.forename: Sana N
authors.initials: SN
authors.lastname: Khan
authors.affiliation: Department of Obstetrics and Gynecology, the C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, MI 48201, United States of America.
authors.forename: Mili
authors.initials: M
authors.lastname: Thakur
authors.affiliation: Department of Obstetrics and Gynecology, the C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, MI 48201, United States of America.
authors.forename: Hamid-Reza
authors.initials: HR
authors.lastname: Kohan-Ghadr
authors.affiliation: Department of Obstetrics and Gynecology, the C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, MI 48201, United States of America.
authors.forename: Sascha
authors.initials: S
authors.lastname: Drewlo
authors.affiliation: Department of Obstetrics and Gynecology, the C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, MI 48201, United States of America.
authors.forename: Ghassan M
authors.initials: GM
authors.lastname: Saed
authors.affiliation: Division of Nephrology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, United States of America.
authors.forename: Subramaniam
authors.initials: S
authors.lastname: Pennathur
authors.affiliation: Department of Obstetrics and Gynecology, the C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, MI 48201, United States of America.|Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, United States of America.
authors.forename: Husam M
authors.initials: HM
authors.lastname: Abu-Soud
mesh_terms: D000818:Animals; D018920:Coculture Techniques; D054885:Cumulus Cells / Q000166:cytology; D008262:Macrophage Activation; D017737:Macrophages, Peritoneal / Q000201:enzymology / Q000276:immunology*; D008677:Metaphase; D051379:Mice; D009865:Oocytes / Q000166:cytology*; D009195:Peroxidase / Q000378:metabolism*
chemical_list: D009195:Peroxidase