The Effect Of Orexin A On The Proliferation, Maturation And Function Of Ovarian Granulose Cells In Sheep

As an important neuropeptide in the hypothalamic, Orexins can project onto the the genital organs and brain regions Widely to regulate the basic activities. Its receptor can be expressed in the reproductive system. Through the axis of hypothalamus-pituitary’ gonadal, hypothalamic orexins and ovarian E2 have mutual influence and feedback regulation between positive and negative to adjust reproductive function. Ovarian granulosa cells play an important role in the formation of follicles and corpus. In addition, it has the function of maintaining ovary and corpus by secreting gonadal hormones. Therefore, explore the role of orexins in the process of granulosa cells growth has theoretical significance on mammalian reproductive function research.This experiment was to culture sheep ovarian granulosa cells in vitro. Add four concentrations orexins of 0.05,0.2,0.5,1 ug/mL after Luteinizing for 0,2,6,12,24 and 48 h. Using qPCR, Western Blot and confocal laser scanning technology to know the influence, and study the effect of orexins A on the growth and development of the granulosa cells. The results were as follows:1.Stimulate luteinizing granulosa cells, Orexins A couid regulate CYP11, StAR and 3 beta HSD mRNA expression. Add four dose group of orexins A, CYP11, StAR and 3β-HSD gene expression had obvious upward trend as the extension of time; Orexins A CYP11, StAR and 3β-HSD gene expression was higher than other group at 24 h and 48 h; Orexins increased significantly at 48 h.2. Orexins A stimulated luteinizing granulosa cells, which could stimulate the expression of P-AMPK and P-ERK1/2 protein effectively after adding orexins A 2 h and 6.h. P-AMPK expression were significantly increased in 0.05 μg/mL and 0.1 μg/mL dose groups. The same dose group decreased gradually from 12h to 24h. The P-ERK expression of 0.05 μg/mL and 0.1μg/mL groups were significantly increased by the addition of Orexins A.0.05 μg/mL and 0.1 g/mL groups reached the platform stage from 12 h to 24 h.0.1 μg/mL group increased significantly after the stage of the platform stage. It showed that the orexins A has positive role in regulating the growth and development of luteinizing granulosa cells.3. Orexins A stimulated luteinizing granulosa cells has significant control effect on the expression of CYP11, StAR and 3 beta HSD protein. The results of CYP17, StAR and 3β-HSD protein expression were consistent with the results mRNA expression at 24 h and 48 h.