ERYPTOSIS AS A LINK BETWEEN BACTERIAL REPRODUCTIVE TRACT INFECTION AND ANEMIA IN PREGNANT WOMEN

Abstract

Genital tract infections of various etiologies and anemia remain common in pregnant women. At present, there is a paucity of data on the impact of reproductive tract infections on the development of anemia in pregnant women, with particular emphasis on the contribution of eryptosis. The aim of the current study was to assess the ability of bacterial, viral and fungal reproductive tract infections to trigger eryptosis of circulating erythrocytes in pregnant women.Methods. The study included 47 pregnant women. According to the pathology of the infection, the patients were divided into three groups: 14 women with bacterial, 12 women with viral, 11 women with fungal infections of the reproductive tract and 10 pregnant women without extragenital and obstetric pathology (control subjects). The following methods were used to determine infection: bacteriologic, microscopic, polymerase chain reaction, immunoenzymatic, etc. Flow cytometry- based detection of eryptosis parameters (cell morphology, membrane phospholipid scrambling, and oxidative stress) was performed in pregnant women with reproductive tract infections of bacterial, viral, or fungal origin and in healthy individuals. Annexin V-FITC staining and 2’,7’-dichlorodihydrofl uorescein diacetate (H2DCFDA) staining were used to quantify phosphatidylserine exposure and reactive oxygen species (ROS) generation, respectively. Fluorescence was acquired using a BD FACSCanto™ II fl ow cytometer. ANOVA test was used to compare numerical data of eryptosis assays. Post-hoc Bonferroni test was applied afterwards. Data were processed by Graph Pad Prism 5.0 software (USA). The diff erence was considered statistically signifi cant at p < 0.05. The research was conducted in accordance with the fundamental principles of Good Clinical Practice (GCP, 1996), the Council of Europe Convention on Human Rights and Biomedicine (April 4, 1997), the Helsinki Declaration of the World Medical Association on Ethical Principles for Medical Research Involving Human Subjects (1964-2008), as well as the Ministry of Health of Ukraine Order No. 690 dated September 23, 2009 (amended by the Ministry of Health of Ukraine Order No. 523 dated July 12, 2012). The research design was discussed and approved at the meeting of the Medical and Ethical Committee of the Kharkiv National Medical University (Protocol No. 19 dated May 19, 2024). The paper is an excerpt from the scientifi c research project of the Department of Obstetrics and Gynecology No. 2 at Kharkiv National Medical University, entitled «Optimisation of diagnostics, treatment of diseases of the reproductive system and pregnancy complications in women with extragenital pathology» (state registration number 0121U110923).)Results. Erythropoiesis parameters were evaluated in pregnant women with bacterial, viral and fungal reproductive tract infections. Pregnancy- associated bacterial reproductive tract infections were associated with increased phosphatidylserine externalization in circulating erythrocytes, without ROS accumulation or cell shrinkage. Erythrocytes from patients with genital tract infections of viral or fungal origin show no signs of eryptosis, such as cell membrane scrambling, oxidative stress, or changes in cell morphology.Conclusions. Reproductive tract infections of bacterial origin in pregnant women are associated with accelerated eryptosis, which may contribute to anemia in these women. Exclusively bacterial genital tract infection was shown to induce lipid membrane scrambling and hence eryptosis in erythrocytes from pregnant women. Phosphatidylserine externalization did not diff er signifi cantly between women with viral or fungal infections and healthy controls. Eryptosis induced by bacterial genital tract infection was not associated with cell shrinkage or ROS overproduction.