PATHOGENETIC JUSTIFICATION OF THE USE OF PROBIOTICS IN PREPARATION PROGRAMS FOR ASSISTED REPRODUCTIVE TECHNOLOGIES IN WOMEN WITH ENDOMETRIOSIS BASED ON STUDYING INDICATORS OF THE LEVEL OF MESOTHELIN, mRNA IL1β, HMGB1, NLRP3-INFLAMMASOME
Authors: Bakun Oksana , Koval Halyna , Muzyka Natalya , DuBuske Lawrence , Slobodian Ksenia 
Published: January 2025

Introduction. Treatment of infertility in women with endometriosis is an important and unsolved problem both in Ukraine and in the world. Along with this, in this cohort of patients, the efficiency of using assisted reproductive technologies (ART) remains low. The optimal approach in the program of preparation for ART of women with endometriosis is precisely probiotics with the content of lactobacilli as potential factors for reducing the production of provoking mediators of endometriosis and increasing the chances of a successful pregnancy.
Aim: To justify the feasibility of including probiotics in the program of preparation for intrauterine reproductive technologies in women with endometriosis-associated infertility.
Methods and materials. The study included 88 women with endometriosis associated with infertility (the main group) and 68 women with tubal infertility (the control group). To analyze the expression of the NLPP3-inflammasome gene, HMGB1, IL1β and determine the relative normalized expression of NLRP3, HMGB1, IL1β mRNA, we used the polymerase chain reaction with reverse transcription in real-time (RT-PCR). The mesothelin level in blood serum was determined using the solid-phase enzyme-linked immunosorbent assay (ELISA).
Results. In earlier stages of the work, the levels of pro-inflammatory markers such as the peritoneal damage marker mesothelin and the expression of NLRP3, IL-1β and HMGB1 in serum and locally in the peritoneal fluid were investigated in women with endometriosis associated with infertility, and results were obtained indicating a probable increase all the indicators listed above. In particular, a significantly higher average level of mesothelin concentration in blood plasma was observed in the main group compared to the control group (0,73±0,01 ng/ml and 0,49±0,01 ng/ml) (p<0,05); NLRP3-inflammasome mRNA expression level (44,43±3,72 c.u. and 0,85±0,03 c.u.) (p˂0,001); IL1β mRNA gene expression (26,47±0,01 c.u. and 0,45±0,01 c.u.) (p˂0,001) and HMGB1 mRNA expression (11,91±0,01 c.u. and 1,00±0,01 c.u.) (р˂0,001). When evaluating these parameters depending on the nature of infertility (primary or secondary), results were also obtained, which indicate that women with endometriosis and primary infertility had significantly higher levels of mesothelin, NLRP3, IL-1β, and HMGB1 inflammasome activity. In particular, the level of mesothelin in women of the main group with primary infertility was significantly higher compared to women with secondary infertility both in blood serum and peritoneal fluid, respectively (0,86±0,001 ng/ml and 0,69±0,001 ng/ ml) (p<0,01) to (0,67±0,001 ng/ml and 0,42±0,001 ng/ml) (p<0,001).
Conclusions.

The study confirmed that the mRNA expression levels of NLRP3, IL-1β, HMGB1 and mesothelin correlate with the severity of endometriosis and can be potential markers of disease progression. These molecules are promising targets for further therapeutic strategies in the treatment of infertility-associated endometriosis.
The study showed that the level of mesothelin in blood serum is 1,25 times higher in women with primary infertility than with secondary infertility and correlates with the severity of the disease, which indicates its possibility to be an independent marker for the diagnosis of endometriosis.
The literature data indicates that the use of probiotics in programs of preparation for assisted reproductive technologies (ART) in women with endometriosis is appropriate.