{"paper_id":"020e6cb8-ac28-4488-a3ae-d25e1de14205","body_text":"ASSESSMENT OF EXPRESSION LEVELS OF NLRP3 INFLAMMASOMES, IL-1β, HMGB1 mRNA IN THE WHOLE BLOOD OF WOMEN WITH ENDOMETRIOSIS ASSOCIATED WITH INFERTILITY\nAbstract\nIntroduction. As it is known, endometriosis is a chronic process that is accompanied by chronic pel- vic pain. Currently, there is an on-going discussion about non-invasive diagnostic markers. But there is still no single specific marker. The pathogenesis of chronic inflammation involves a progressive course, which is accompanied by long-term maintenance of the inflammasome in an active state due to excessive accumulation of intracellular and then extracellular DAMPs, their inefficient utilization by autophagy and stress-reducing cellular systems, local production of pro-inflammatory cytokines IL-1β, IL-18, and as well as the accumulation of M2 macrophages and helper T-lymphocytes of the 2nd type, tissue damage and the development of fibrosis.\nHMGB1 is present in almost all cell types. It can also be released extracellularly, where it mediates the activation of innate immune responses, including chemotaxis and cytokine release. HMGB1 was origi- nally described as a protein that binds nuclear DNA. It is evolutionarily highly conserved and functions as a nuclear cofactor in transcriptional regulation.\nRegulation of IL-1 activity is carried out at two levels and requires the action of two stimulating signals – the first causes the expression of the inactive form of interleukins, proIL-1α and proIL-1β, in which the ac- tive interleukin is, connected to an additional peptide chain, which completely blocks the activity of proIL-1β and significantly to an extent – proIL-1α. The second level of regulation is carried out at the stage of activa- tion of prointerleukins by proteolytic cleavage of the propeptide by the intracellular protease caspase-1, which is synthesized in the form of an inactive proen- zyme.\nThe aim of the study. To study the gene expres- sion levels of NLRP3-inflammasome mRNA, IL-1β, HMGB1 in the whole blood of women with endometriosis associated with infertility and to reveal their role in the development of the disease.\nMaterials and methods. To analyze the expres- sion of NLRP3-inflammasome, IL-1β, HMGB1 genes and to determine the relative normalized expression of NLRP3, IL-1β, HMGB1 mRNA, we used the poly- merase chain reaction with real-time reverse transcription (RT-PCR) method in 50 women: 40 women with endometriosis and 10 women with tubal infertility. The object for molecular genetic studies by the RT- PCR method was the fraction of mononuclear cells isolated from the whole blood of patients with endo- metriosis.\nStatistical processing of the results of this sec- tion was carried out using non-parametric methods, namely the Wilcoxon-Mann-Whitney test, in which the indicators were considered reliable at p<0.05.\nResults. On the basis of the conducted study, it was established that the level of NLRP3-inflamma- some mRNA expression in the blood of women with endometriosis associated with infertility is signifi- cantly increased compared to the indicators of the control group (44.43±3.72 c.u. and 0.85±0.03 c.u.) (р<0.001). Such results of relative normalized expres- sion of the NLRP3 gene turned out to be quite unexpected. It is obvious that such a high activation of the receptor is associated with the presence of potential ligands capable of activating it. Among the known objects with such ability and presence in the reproductive tract (endometrium) of women, a number of obligate and conditionally pathogenic microorganisms can be distinguished, which contain antigenic configurations (lipoproteins, peptidoglycan, glycolipids, zymosan of mushrooms) capable of activating NLRP3. However, opportunistic pathogens are not subject to manda- tory drug elimination. However, it is not excluded that this very fact supports the growth of endometrioid ectopies with the help of mechanisms that are not fully understood. On the basis of our research, it was es- tablished that the expression of IL-1β mRNA gene in the blood of women with endometriosis associated with infertility is significantly increased. At the same time, a significant difference with the control group was established (26.47±0.01 c.u. and 0.45±0.01 c.u.) (р 0.001).\nThe range of all obtained values of the relative normalized amount of mRNA of the IL-1β gene was 0.004-153.01 (median – 26.78). On the basis of our study, it was established that the expression of HMGB1 mRNA in the blood of women with endome- triosis associated with infertility is probably increased compared to the control group (11.91±0.01 c.u. and 1.00±0.01 c.u.) (p 0.001). Thus, in the blood samples of 95% of women, the expression level of the HMGB1 mRNA gene is significantly higher compared to the control, with a preference for high values (p<0.001).\nConclusions. The level of NLRP3-inflamma- some mRNA expression in the blood of women with endometriosis associated with infertility is signifi- cantly increased compared to the indicators of the control group (44.43±3.72 c.u. and 0.85±0.03 c.u.) (p<0.001). It was established that the expression of the IL-1β mRNA gene in the blood of women with endometriosis associated with infertility was signifi- cantly increased compared to the indicators of the control group (26.47±0.01 c.u. and 0.45±0.01 c.u.) (p<0.001). The expression of HMGB1 mRNA in the blood of women with endometriosis associated with infertility is significantly higher (11.91±0.01 the main group and 1.00±0.01 of the control group) (p<0.001). The study found that endometriosis associated with infertility is a chronic inflammatory disease, HMGB1 has a slight pro-inflammatory activity and forms com- plexes with IL-1β, and thus enhances its biological activity and thus the expression of the Nlrp3 inflam- masome. Thus, increased expression of mRNA genes NLRP3-inflammasome, IL-1β, HMGB1 in endometrio- sis can be considered as a protective reaction.\nDownloads\nReferences\nCardoso JV, Machado DE, da Silva MC, de Mello MP, Berardo PT, Medeiros R, Perini JA. 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