THE IMPACT OF PREVENTIVE TREATMENT IN PREGNANT WOMEN AT HIGH RISK OF PREECLAMPSIA ON THE IMBALANCE OF PRO- AND ANTI-INFLAMMATORY CYTOKINES, INOS, AND ARGINASE ACTIVITY

Preeclampsia complicates 7–10% of pregnancies and ranks as the third to fourth leading cause of maternal mortality. An imbalance in cytokine production, characterized by a predominance of pro-inflammatory cytokines, leads to systemic endothelial dysfunction. This dysfunction results in inadequate trophoblast invasion into the spiral arteries of the uterus, contributing to various gestational complications. Aim: To investigate the levels of pro- and anti-inflammatory cytokines, as well as the activity of inducible NO synthase (iNOS) and arginase, in the cervical mucus of pregnant women at high risk of preeclampsia. Additionally, to evaluate the effectiveness of preventive treatment in restoring cytokine balance and normalizing nitric oxide metabolism. Materials and methods: The study included 110 pregnant women, divided into a control group (CG) of 28 healthy pregnant women and a main group (MG) of 82 women at high risk of preeclampsia. The high-risk status was determined based on anamnestic factors and reduced blood flow in the placental spiral arteries at 18–20+6 weeks of gestation. Within the main group, 33 women did not receive advanced secondary prevention (MG-I), while 49 women (MG-II) were treated with metformin, vitamin D3, and quercetin with povidone. Cervical mucus samples were collected using a sterile brush, suspended in 2 mL of 0.2 M Tris-buffered solution (pH = 7.4), and stored frozen. Levels of pro- and anti-inflammatory cytokines (IFN-γ, TNF-α, IL-10) were measured using standard commercial reagent kits. iNOS activity was determined by subtracting constitutive NO synthase (cNOS) activity from total NO synthase activity. Arginase activity was assessed by measuring the difference in L-ornithine concentrations before and after incubation at 37.0°C. Results were expressed in µmol/min per 1 g of cervical mucus protein. The study adhered to biomedical ethics guidelines. Statistical analysis was performed using the “STATISTICA” software (“StatSoft Inc.,” USA), including calculation of mean values (M), variance (σ), standard errors (m), and Student’s t-test. This study is part of an initiative research project conducted by the Department of Obstetrics and Gynecology No. 2 at Poltava State Medical University, titled “Optimization of Approaches to Pregnancy Management in Women at High Risk of Obstetric and Perinatal Pathology” (State Registration No. 0122U201228, implementation period 2022–2027). Results and Discussion: In the high-risk group without preventive treatment (MG-I), elevated levels of pro-inflammatory cytokines and reduced levels of the anti-inflammatory cytokine IL-10 were observed in cervical mucus. iNOS activity was significantly increased, while arginase activity was decreased compared to the control group. Significant correlations were identified between iNOS activity and the levels of TNF-α (r = 0.77; P < 0.001) and IFN-γ (r = 0.55; P < 0.01). A positive correlation was found between decreased IL-10 levels and reduced arginase activity (r = 0.56; P < 0.02), while a negative correlation was observed between increased iNOS activity and decreased arginase activity (r = -0.43; P < 0.05). In the treatment group (MG-II), preventive therapy reduced pro-inflammatory cytokine levels: IFN-γ decreased by 1.2 times and TNF-α by 1.8 times compared to MG-I. Conversely, IL-10 levels increased by 1.5 times. iNOS activity decreased by 1.9 times, while arginase activity increased by 1.6 times. Clinically, preventive treatment reduced the incidence of preeclampsia by 1.5 times and perinatal mortality by 1.8 times. Conclusions: Pregnant women at high risk of preeclampsia who did not receive preventive treatment exhibited elevated levels of pro-inflammatory cytokines (IFN-γ and TNF-α) and reduced levels of the anti-inflammatory cytokine IL-10 in cervical mucus at 28–34 weeks of gestation. Additionally, iNOS activity was increased, while arginase activity was decreased. In contrast, women who underwent preventive treatment (MG-II) demonstrated reduced pro-inflammatory cytokine levels, increased IL-10 levels, decreased iNOS activity, and increased arginase activity. Clinically, preventive treatment reduced the incidence of preeclampsia by 1.5 times and perinatal mortality by 1.8 times.