The effects of Salvia officinalis L.   on granulosa cells and in vitro maturation of oocytes in mice

Background: Salvia officinalis L. has been used since ancient times but there are little data about effects of this herb on normal reproductive cells.
Objective: To investigate the toxicity effects of Salvia officinalis L. on granulosa cells (GCs) and maturation of oocytes.
Materials and Methods: GCs and oocytes were extracted from superovulated ovaries of immature mice. The cells were treated with concentrations of 10, 50, 100, 500, and 1000 μg/ml of Salvia officinalis hydroalcoholic extracts and compared with the control culture. Bioviability, chromatin condensation, estradiol and progesterone concentrations, lipid synthesis, apoptosis, and alkaline phosphatase activity of GCs were measured. In vitro maturation of oocytes by determination of different maturation stages of oocytes including germinal vesicle, germinal vesicle breaks down, and metaphase II were examined.
Results: The results revealed that 500 and 1000 μg/ml concentrations of Salvia officinalis L. were toxic. The most of the GCs were in the early stages of apoptosis in 100 μg/ml treated culture and cell death happened with 500 μg/ml treatment. Progesterone concentration was reduced in 100 μg/ml and higher doses but estradiol concentration and alkaline phosphatase showed opposite effects. The lipid droplets content of GCs reduced significantly in all groups especially in 500 and 1000 μg/ml. Finally, oocyte’s nucleus and cytoplasm showed a high level of condensation, and meiosis rate reduced in all treated cultures.
Conclusion: Our findings suggested that higher dose of Salvia officinalis hydroalcoholic extracts inhibits, oocyte maturation, GCs bioviability, proliferation, and secretion.

Keywords
Granulosa cell; In vitro maturation; Oocytes; Ovary; Salvia officinalis.