Study On The Effect Of Orexin A On Progesterone Secretion By Ovarian Luteinized Granulosa Cells In Sheep By ABCG1 Protein

Orexin A is a hormone secreted by the hypothalamus,which is widely involv ed in life activities and material metabolism by binding to G protein-coupled rece ptors.In the early stage,our laboratory concluded that progesterone secretion incr eased after treatment with sheep ovary luteinized granulosa cells at a concentratio n of 10 nmol/L Orexin A for 24 hours.This experiment is based on the previous conclusions.Orexin A-added sheep ovary luteinized granulosa cells were sequenc ed and analyzed by transcriptome,and the ABCG1 gene with significantly decreas ed expression was obtained.Because ABCG1 is a cholesterol reverse transport pro tein,it can maintain the homeostasis of cholesterol in cells.Cholesterol,as a raw material for progesterone synthesis,affects cholesterol transport and may directly affect progesterone synthesis and secretion.Therefore,this experiment explored the connection between Orexin A,ABCG1 and progesterone.The main result was that the addition of Orexin A to the luteinized granulosa cells and the control group were analyzed by transcriptome sequencing.There we re genes with significantly different expression levels,including the down-regulated ABCG1.The results of q-PCR verification confirmed that the expression of ABC G1 gene in luteinized granule cells after the addition of Orexin A was significantl y different,which was consistent with the sequencing results;ELISA results show ed that the progesterone secretion increased after the luteinized granule cells were treated with different concentrations of ABCG1 inhibitors And the effect is best when the concentration is 20 nmol/L.Progesterone secretion increased after the ad dition of ABCG1 inhibitor and Orexin A in luteinized granulosa cells,indicating t hat inhibition of ABCG1 can affect the secretion of fine progesterone in luteinize d granules added with Orexin A;Western blot results showed that ABCG1 in lute inized granule cells after adding Orexin A Expression decreased,StAR expression increased;ABCG1 expression decreased after luteinized granulosa cells added ABC G1 inhibitor group,StAR expression increased;ABCG1 expression decreased after luteinized granule cells added ABCG1 inhibitor and Orexin A,and StAR expressi on level rise.The results were confirmed by q-PCR and ELISA results,indicating that the inhibition of ABCG1 can affect the expression levels of ABCG1 and St AR in luteinized granulosa cells and luteinized granulosa cells added with Orexin A.This experiment shows that Orexin A can affect the secretion of progesterone by luteinized granulosa cells of sheep ovary through ABCG1 protein,and inhibiti ng ABCG1 can promote Orexin A to affect the secretion of progesterone by lutei nized granulosa cells of sheep ovary.