Research On Genes Expression Of FoxO1 Regulating Fatty Acid Metabolism In Goose Granulosa Cells

Female ovarian follicle development is closely related to reproductive performance,granulosa cells,the interaction between theca cellsand oocytes,follicles start to complete the development,and maturation.In mammals and birdshas been found ovarian granular cells exist the ability of fatty acids from scratch.The study found that mammalian ovary fatty acid oxidation can be used to power the oocyte development,mature,this in order to ensure the development of follicles and mature.Prophase study did have follicular lipid synthesis ability,the follicle granulosa cells regulation of lipid metabolism or mechanism is not clear,granulosa cells lipid metabolism is still in the exploration stage,This study Tianfu mother goose egg peak as the research object.Detection follicles at different developmental stages fatty acid metabolism-related genes(FoxO1?CPT-1?Apob?PPAR ??PPAR??PPAR ?)mRNA expression levels.FoxO1 as an important transcription factors,cell metabolism by building FoxO1 interference carrier,detection of fatty acid metabolism related genes mRNA,intracellular lipid droplets content changes.To explore the interference within the granulosa cells after FoxO1 gene expression related to fatty acid synthesis,oxidation,research FoxO1 role in granular cell lipid metabolism.On the other hand,adding different concentration(25 ?M,100?M and 150?M)of mildronate,the detection of fatty acid metabolism related genes mRNA,intracellular lipid droplets content changes and related enzyme gene level.To further explore the mildronate effect on fatty acid metabolism in granulosa cells and FoxO1 transcription factor of fatty acid metabolism in granulosa cells related gene expression research.The main results were as follows:(1)The expression of lipid metabolism related genes(FoxO1?CPT-1?Apob?PPAR ??PPAR ??PPAR ?)in the developmental stages of the healthy follicle tissue of the goose(in the separation of the granulosa cellsandthe theca cells)was observed by the qPCR test.The expression ofFoxO1?CPT-1?Apob?PPAR ??PPAR ?andPPAR?were different in different cells and at different developmental stages.(2)Construction of four FoxO1 carrier interference,The interference efficiency of F1393 interference sequence was 67.71%after transfection of granulosa cells,and the interference efficiency was the highest.(3)After the interference FoxO1,compared with control group showed;significantly reduced FAS,ACC?,PPAR?,CPT-1,DGAT2 mRNA expression(P<0.05),increased PPAR?,PPAR?,DGAT1 mRNA expression,(P<0.05),of The mRNA expression of SREBP1 did not change significantly,and the content of TG in granulosa cells increased.(4)With different concentrations(25?M,100?M and 150?M)of mildronate treated granulosa cells,results showed that:compared with the control group,three concentrations mildronate promote the expression of FAS,ACC?,PPAR?,PPAR?,CPT-1,ELOVL6andDGAT2 gene,The mRNA expression of DGAT1 was decreased by 150?M,The decreased the activity of FAS and decreased the content of TG in the cells.In summary,the expression of FoxO1?CPT-1?Apob?PPAR??PPAR? and PPAR y was found in follicular tissue.FoxO1 silence resulted in decreased expression of FAS,ACC?,PPAR?,CPT-1.The expression of FoxO1 gene was promoted by rice hydrazine in the treatment of granulosa cells,and the expression of FAS,ACCa,PPARa,CPT-1 and so on was also increased.