THE EFFECTS OF LIQUIRITIGEN ON THE LUKEMIA INHIBITORY FACTOR RECEPTOR IN GRANULOSA CELLS OF INFERTILE PATIENTS

Abstract

Objectives: Granulosa cells have an important role in oocyte nutrition and maturation, oocyte-sperm interaction events such as capacitation, and acrosome reaction. Therefore, studies with granulosa cells are also important for the healthy development of the oocyte and fertilization. The aim of this study was to investigate the effects of liquiritigenin (LQ) in vitro on granulosa cells that were obtained from the oocyte-cumulus complex of patients who were treated in the in vitro fertilization (IVF) unit due to ovulatory factor (OF, infertile) and male factor (MF, healthy). Materials and Methods: This study included patients who were treated in the IVF unit of the Istanbul University Faculty of Medicine due to OF and MF. Granulosa cells obtained from 5 MF and 5 OF patients were divided into MF, MF-Dimethyl sulfoxide (DMSO), MF-LQ, OF, OF-DMSO and OF-LQ groups in culture medium. After the application, the cells were stained with the leukemia inhibitory factor receptor (LIFR) immunocytochemical staining and the staining intensity was examined with the Aperio ImageScope analysis software and cell morphology. Results: The LIFR immunocytochemistry staining intensity in the granulosa cells of the OF and OF-DMSO groups was significantly decreased when compared to the MF and MF-DMSO groups. The LIFR staining intensity in the granulosa cells of the MF-LQ and OF-LQ groups were determined to be significantly higher than the other groups. In the granulosa cells of OF and OF-DMSO groups, the findings reflected apoptotic cell morphology, but showed no or less apoptotic cell morphology in the MF-LQ and OF-LQ groups. Conclusions: In granulosa cells of LQ-groups, LIFR immunocytochemistry staining intensity algorithms were obtained as significantly higher than in the especially the OF and OF-DMSO groups, and showed the positive effects on apoptotic cell morphology. These results suggest that LQ may be used as an alternative medium agent in IVF units. Therefore, more comprehensive studies are needed for the clinical use of LQ. 

Keywords

• Infertility
• Cumulus-granulosa cells
• Leukemia inhibitory factor receptor

İNFERTİL HASTALARIN GRANÜLOZA HÜCRELERİNDE LİQUİRİTİGENİN’İN LÖSEMİ İNHİBİTÖR FAKTÖR RESEPTÖRÜ ÜZERİNE ETKİLERİ

Abstract

Amaç: Granüloza hücreleri oosit beslenmesi ve olgunlaşması, kapasitasyon ve akrozom reaksiyonu gibi oosit-sperm etkileşimi olayları üzerine önemli bir role sahiptir. Bu nedenle, granüloza hücreleri ile ilgili çalışmalar, oositin sağlıklı gelişmesi ve fertilizasyon için de önemlidir. Çalışmada ovulatuar faktör (OF, infertil) ve erkek faktörü nedeniyle (EF, sağlıklı bireyler) in vitro fertilizasyon (IVF) ünitesine başvuran hastaların oosit-kümülüs kompleksinden elde edilen granüloza hücrelerinde, in vitro ortamda liquiritigenin ’in (LQ) etkilerinin immünositokimyasal olarak incelenmesi amaçlanmıştır. Gereç ve Yöntem: Çalışmada OF ve EF nedeni ile İstanbul Üniversitesi Tıp Fakültesi tüp bebek IVF ünitesine başvuran hastalar alındı. 5 EF ve 5 OF hastasından alınan granüloza hücreleri kültür ortamında EF, EF- Dimetil sülfoksit (DMSO), EF-LQ, OF, OF-DMSO ve OF-LQ gruplarına ayrıldı. Hücreler uygulama sonrası lösemi inhibitör faktör reseptör (LIFR) immünositokimyasal boyaması yapılarak Aperio ImageScope analiz proğramıyla boyanma şiddeti ve hücre morfolojisi bakımından incelendi. Bulgular: OF ve OF-DMSO gruplarının granüloza hücrelerinde LIFR immünositokimya boyanma şiddeti, EF ve EF-DMSO gruplarına göre anlamlı olarak azalmıştır. EF-LQ ve OF-LQ gruplarının, granüloza hücrelerinde LIFR immünositokimya boyanma şiddeti bakımından diğer gruplarla karşılaştırıldığında aralarında anlamlı bir artış olduğu saptanmıştır. OF ve OF-DMSO gruplarının granüloza hücrelerinde, apoptotik hücre morfolojisini yansıtan bulgular gözlenirken, EF-LQ ve OF-LQ gruplarında ise hiç veya daha az olarak gözlenmiştir. Sonuç: LQ-gruplarının granüloza hücrelerinde LIFR immünositokimya boyanma şiddeti algoritmaları, özellikle OF ve OF-DMSO gruplarına göre yüksek saptanmış ve apoptotik hücre morfolojisi üzerine olumlu etkileri gözlenmiştir. Bu sonuçlar, LQ ‘nun IVF ünitelerinde alternatif bir medyum ajanı olarak kullanılabileceğini düşündürmektedir. Bu nedenle LQ ‘nun klinikde kullanılabilmesiyle ilgili daha kapsamlı çalışmalara ihtiyaç duyulmaktadır.

Keywords

• İnfertilite
• Kümülüs-granüloza hücreleri
• Lösemi inhibitör faktör reseptör
• Liquiritigenin
• Infertility
• Cumulus-granulosa cells
• Leukemia inhibitory factor receptor

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