EFFECTIVENESS OF THYROXINE IN SWIM UP SPERM PREPARATION TECHNIQUES IN RELEVANCE TO PHOSPHATIDYLSERINE LEVEL IN IRAQI INFERTILE MEN
Background: Non-genomic actions of thyroid hormones are self-determining of nuclear receptors. They have been studied at the plasma membrane and in the cytoplasm. They involve changes in solute transport, the activity of several kinases, and the translation of certain mRNAs. Also, they affect the control of actin polymerization, which changes the way the cytoskeleton moves and change how the mitochondria breathe. "Direct and indirect swim-up" is a method for preparing sperm. By using the "swim up" method, the sperm were chosen based on how quickly they moved and whether or not they could swim out of the seminal plasma. Phosphatidylserine (PS) one of the major structural phospholipids in mammalian membranes. It is exposed on the head of healthy, moving sperm, and the amount of PS that is exposed grows as the sperm moves through the epididymis. Phosphatidylserine PS receptors on the oocytes are expressed on the microvilli to help sperm:egg fusion.
Aim of study:To assess the non-genomic action of T4 hormone on sperm parameters and its probable use in vitro to improve sperm recovery after an enrichment method like “swim-up” and make a comparison with the conventional swim up techniques. To detect phophotidylserine PS in seminal plasma as a biomarker for competent sperm and make comparison between PS level before and after addition of Thyroxine T4 in both direct and indirect swim up techniques.
Patients and Methods: In this interventional comparative study, 40 infertile men were included during their presence to the infertility clinic at High Institute of Infertility Diagnosis and Assissted Reprductive Technologies,Al-Nahrain University from November 2022 to May 2023. Normozoospermic or mild asthenozoospermic semen samples were used to choose the cases, according to WHO manual standards (1999 -2010). The collected semen samples obtained were divided into five parts: The first part prepared was in vitro sperm characterization before activation, the second part was after activation using direct swim up technique,the third part was after activation using indirect swim up technique while the fourth and fifth parts were after activation using direct and indirect swim up after adding thyroxine T4. A comparison was made between groups for semen paramers. The T4 dose used in this study were selected on the basis of that many doses in different pilot studies were used and found this dose, which is little bit low,is the best dose and used previously in researches for the same purpose. In addition ,measurement of phosphotidylserine levels in the seminal plasma was done .A test called an Enzyme-Linked Immunosorbent Assay was used to find out how much PS there was.
Results: The study investigated the effect of addition of thyroxine 200 μg/ml on semen parameters.Thyroxine causes improvement of sperms motility and morphology and significantly higher results were found from adding thyroxine to direct swim up media. Seminal fluids phosphatidylserine levels were increased after the four methods (direct and indirect swim up with and without thyroxine) and significantly higher in direct swim up activation technique with thyroxine than before activation.
Conclusion: Thyroxine was found to have stimulatory effect on sperm motility percentage, when the semen was treated with 200 μg/ml concentration of Thyroxine by direct swim up technique. Thyroxine application increased the recovery rate of morphologically normal sperm when direct and indirect swim up techniques were used. The rise in seminal fluid phosphatidylserine levels and PS externalization in human sperm is used as an indicator for the estimation of sperm quality. From the statements above, a new physiological appliance was suggested to artificially develop insemination. The discussion opens windows to explore indefinite paths involved in sperm capacitation and gives resourceful opinions to better understand infertility mechanisms.
Key Words: Thyroxine, PhosphotidylSerine (PS), Direct and indirect Swim Up, Infertile men, sperm motility.