Immunohistochemical and electron microscopic investigation of the effects of liquritigenin on granulosa cells of infertile patients with ovulatory factor

Infertility is a global health problem that has been increasing day by day. The fact that granulosa cells (GCs) have an important role in the oocyte nutrition and maturation as well as the oocyte-sperm interaction makes them appear as an option in the diagnosis and treatment of infertility. Liquiritigenin (LQ) is a flavonoid compound derived from Glycyrrhiza radix. This experimental study aimed to examine the effects of LQ on GCs obtained from patients treated in an in vitro fertilization (IVF) unit due to the ovulatory factor (OF, infertile) and male factor (MF, healthy individuals). GCs obtained from patients were divided into groups as MF, MF Dimethyl sulfoxide (DMSO), MF-LQ, OF, OF-DMSO and OF-LQ. LQ (40 μM) and DMSO (5 μL) were applied to culture mediums of determined groups for 24 hours. SIRT2 staining intensity and cell morphology were evaluated by immunocytochemical staining. Ultrastructural morphologies of GCs were examined by transmission electron microscopy. The intensity of SIRT2 immunoreactivity in GCs of OF and OF-DMSO groups was significantly decreased compared to those of MF and MF-DMSO groups. SIRT2 immunoreactivity increased significantly in EF-LQ and OF-LQ groups when compared to other groups. In OF and OF-DMSO groups, ultrastructural findings reflected apoptotic cell morphology but MF-LQ and OF-LQ groups showed less apoptotic cell morphology These results demonstrate that LQ has positive in vitro effects on morphology and SIRT expression in GCs, suggesting that LQ can be used as an alternative medium agent in IVF procedures. However, further detailed studies are needed.