Research Of Melatonin On Bovine Theca Cells Apoptosis And Steroidogenesis

The apoptosis and steroidogenesis of Theca cells is indispensable in follicular development and atresia.Besides,the specific mechanism of the effect of melatonin on steroidogenesis and apoptosis needs further study.Therefore,the study of melatonin on the steroidogenesis and apoptosis of thecal cells can further elucidate the role of melatonin in follicular development and atresia.In this study,we used the 3-4 mm bovine follicles as the experimental material and studied the effect of melatonin on theca cells steroidogenesis and apoptosis in vitro,and further elucidated the specific regulatory mechanism.The results are as follows:1.Effects of melatonin on apoptosis and steroidogenesis on bovine theca cellsThe follicles were treated with 0 ng/m L,1 ng/m L,10 ng/m L and 100 ng/m L melatonin.Tearing the follicular intima,the expression of melatonin receptor MT1 and MT2 in follicular intima cells was detected by RT-q PCR.The results of RT-PCR showed that 10 ng / ml melatonin has the most obvious effect.Follicle culture medium was added with 10 ng / ml melatonin.RT-q PCR was used to detect the expression of LHR,the genes related to steroid production and the expression of BCL2,BAX and Caspase.Elisa was used to detect the secretion of androstenedione and progesterone in the follicle culture.The results showed that melatonin significantly promoted the expression of LHR and STAR and significantly promoted the secretion of progesterone.Melatonin significantly promoted the expression of anti-apoptotic gene BCL2,and the ratio of BCL2 and BAX increased.TUNEL staining was used to detect the apoptosis of follicular cells cultured in vitro after 10 ng/m L melatonin treatment.The results showed that the apoptotic rate of melatonin group was lower than that of control group.This study shows that melatonin affects the steroidogenesis and inhibits the apoptosis of bovine theca cells.2.Effects of melatonin on apoptosis and steroidogenesis on theca cells: role of MT1 and MT2 The follicles were treated with 10 ng/m L melatonin,by the addition of 10 μM 4P-PDOT or 10 μM Luzindole to detect the expression of melatonin receptor MT1,MT2 and LHR,STAR and the anti-apoptotic gene BCL2.Elisa was used to detect the secretion of progesterone in the follicle culture.The results of RT-q PCR showed that melatonin significantly promoted the m RNA expression of MT1 and MT2,Luzindole inhibited the expression of MT1 and MT2,and 4P-PDOT inhibited the expression of MT2.Melatonin could significantly promote the expression of LHR,STAR and the anti-apoptotic gene BCL2.Luzindole or 4P-PDOT significantly inhibited the expression of LHR,STAR and BCL2,and significantly inhibited the secretion of progesterone.This study shows that MT1 and MT2 mediate the regulation of melatonin on follicular theca cells apoptosis and steroidogenic.3.Effects of melatonin on apoptosis and steroidogenesis of theca cells: role of PI3K/AKT signaling pathwayThe follicles were treated with 10 ng / m L melatonin,by the addition of 25 μM LY294002 to detect the expression of LHR and STAR and the anti-apoptotic gene BCL2.Elisa was used to detect the secretion of progesterone in the follicle culture.Melatonin could significantly promote the expression of LHR,STAR and the anti-apoptotic gene BCL2.LY294002 significantly inhibited the expression of LHR,STAR and BCL2.The study shows that PI3K/AKT pathway plays a role in melatonin-mediated follicular theca cells apoptosis and steroidogenesis.Conclusion: MT1/MT2 and PI3K/AKT signaling pathway mediates the effect of melatonin on steroidogenesis and apoptosis of theca cells.