P-125: The Effect of Eicosapentaenoic Acid (Omega-3) on The Expression of HAS2 in Cumulus Cells on Mice

Background: Omega-3 fatty acids can modify gene and protein expression through altering the function of transcription factors controlling gene expression. Cumulus expansion, is a critical process for normal oocyte development, ovulation, and fertilization. This event needs to the synthesis of hyaluronic acid and its production is controlled by the enzyme hyaluronic acid synthase 2 (HAS2). The aim of this study was to determine if Omega- 3 can affect on the expression of HAS2 in Cumulus cells and how its outcome in the in vitro fertilization was. Materials and Methods: For this purpose, 30 female mice were super ovulated with hMG and HCG as IP injection. For the collection of cumulus- oocyte complexes(COCs) mice were killed by cervical dislocation and COCs were collected by flashing method. COCs were cultured in 2cc universal medium supplemented with 20 λ omega-3 and incubated for 4-6 hours. Surrounding cumulus cells were removed mechanically by using 20 λ hyaluronidase. The cumulus cells was stored at- 80°C until real-time PCR. PCR was performed by using a master mix containing CYBR green. The expansion level of mRNA HAS2 was evaluated by real-time PCR. For the IVF sperms from male mice were added to the medium drops containing 50 COCs in each group. Results: The result showed that the level of expansion of HAS2 in experimental group comparing with control was significantly increased. Conclusion: The result indicates that omega-3 could improve Cumulus expansion through increasing the amount of expansion of HAS2.