Effect of Red Clover Leaf Extract on Expression of Genes Involved in Follicogenesis and Proliferation of Ovarian Granulosa Cells of NMRI Mice

Subject Areas : Journal of Animal Biology

Farzaneh Rangamiz ¹ , Javad Baharara ² , Khadijeh Shahrokhabadi ³

1 - Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
2 - Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
3 - Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran

Received: 2020-05-11 Accepted : 2020-07-22 Published : 2020-11-21

Keywords: Infertility, Red Clover, Follicogenesis, Granulosa Cell, BMP15 Genes, FOXO1 Genes,

Abstract :

Infertility is one of the major medical problems in today’s world. Today, some studies have addressed the effects of phytoestrogenic compounds in the treatment of infertility. Red clover is one of the herbs containing phytoestrogenic compounds. In the present study, the effect of red clover leaf extract on expression of genes involved in follicogenesis and proliferation of granulosa cells in NMRI mice was investigated. In this experimental in vitro study, 30 NMRI 18-20 immature female rats were used. For experiments, follicular granulosa cells were isolated, and in vitro cells were randomly divided into 6 groups.Control (untreated) and experimental groups with concentrations (10, 15, 20, 40, 80 μg/ml) of treated red clover leaf extract and cell viability were evaluated by trypan blue staining and MTT staining. Expression changes of BMP15 and FOXO1 genes were evaluated by Real Time PCR. The results of trypan blue staining and MTT assay showed that cell viability decreased with increasing concentration and time of treatment with clover leaf extract. Moreover, the expression level of BMP15 and FOXO1 genes was significantly increased in the 10 μg/ml red clover leaf extract compared to the control group (p < 0.05). Due to the effect of red clover leaf extract on granulosa cell proliferation and expression of genes involved in follicogenesis, this plant can be considered as a suitable candidate for clinical studies in infertility treatment.  

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