Fig 1: The presence of NETs (MPO and histones: H2A, H2B, and H3) in the chorion and the placental decidua. Chromogen 3.3'-diaminobenzidine: (1) anti-MPO, (2) anti-H2A, (3) anti-H2B, (4) anti-H3 antibodies; (A) control group, without granulocytes (magnification 200x); (B) women with miscarriage, with granulocytes (magnification 400x); (C) women with miscarriage, with NETs (arrows) (magnification 400x).
Fig 2: Proposed mechanism of NO/NETs-dependent oxidative stress formation in women who had miscarried. NOS-dependent neutrophils-derived NO promotes miscarriage by lipid peroxidation, by NT and MDA. NOX-dependent stimulating ROS production, could affect miscarriage. We hypothesized if NOX-dependent NETs forming is the cause of miscarriage. Based on observed two population of measured placenta and serum markers we suggest two different NETs-involving mechanisms occurrences of miscarriage. In NETs positive patients (increase MPO and PTX3, decrease NO and cfDNA in serum), miscarriages were supported by direct harmful influence of NETs components. Whereas, in NETs negative patients (increase NO and NT, decrease PADI4 and cfDNA, but increase MPO and PTX3 in serum) with very high-level of oxidative stress markers (NO and NT)—dysregulation of oxidative and antioxidant balance, which suggest oxidative stress-dependent impairment and miscarriage. Abbreviations: NETs, neutrophil extracellular traps; NOS, nitric oxide synthase; NOX, NADPH oxidase; ROS, reactive oxygen species; NT, nitrotyrosine; MDA, malondialdehyde; PADI4, peptidylarginine deiminase 4; cfDNA, cyrkulating free DNA; MPO, myeloperoxidase; PTX-3, pentraxin 3; histones 2A, 2B, 3—H2A, H2B, H3.
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