The Study On Effect Of Orexin-a On Progesterone In Sheep Luteinize Granulose Cells

Orexin as a small molecule polypeptide,synthesized by the lateral hypothalamic area.Orexin can regulate the human appetite.It also directly regulates the activity of the animal reproductive endocrine.Previous studies have shown that mammalian production performance is closely related to the content of serum progesterone secreted by ovarian granulosa cells.The function of the animal reproductive system affected by lutein development and secretion of honnones.Therefore,the study of mammalian progesterone secretion is important to regulation of mammalian production performance.In order to observe the effects of different concentrations of OrexinA on progesterone-related proteins in ovarian lutealized granulosa cells of sheep,the study cultured sheep ovarian granulosa cells in vitro,The cells were lutealized,following the incubation with OrexinA in 0h,24h,48h,72h after identification of lutealized treatment,Briefly,ELISA was used to detect the progesterone secretion of sheep ovarian granulosa cells to select the optimal lutealized time.The next experiment added different concentrations(1 nM,10 nM,58 nM,100 nM,145 nM)of the Orexin A incubated in Oh,24h,48h,72h,respectively.Using ELISA technique to detect the progesterone secretion of ovarian granulosa cells in sheep.The expression of key proteins STAR,3 ?-HSD and P450(CYP11)in the process of progesterone secretion in ovarian granulosa cells was detected by Western Blot.This work demonstrated that:1.The lutealized ovarian granulosa cells the highest secretion of progesterone at 24 h.2.The expression of progesterone in ovarian lutealized granulosa cells increased gradually and then decreased after add different concentrations of Orexin A at the same time,and the expression reached the highest at 10nM.The expression of progesterone in ovarian luteinized granulocytes of sheep added the same concentrations of Orexin A incubated in different time was gradually increased and then decreased,and the expression of time reached the highest at 24h.3.Add the different concentrations Orexin A incubated in the same time,the expression of STAR,3 ?-HSD and P450(CYP11)in sheep ovarian granulosa cells were gradually increased and then decreased at different concentrations,and the expression reached the highest at 10nM and 58nM.Add the same concentrations Orexin A incubated in the different time,the expression of STAR and 3 ?-HSD was gradually increased and then decreased in the same concentration,and the expression of time reached the highest at 24h.But the concentration of P450(CYP11)decreased with increasing time.