Eeffcts Of Dianji Recipe On PCOS Rats Ovarian Grnualosa Cell Steoridogenesis And Associated Gene Expression

Research programme:PCOS rats grow in vitro culture and are injected with Serum containing Dianji Recipe.To observe the effect of Dianji Recipe on secretion function of ovary granule cell of PCOS rat, further research for influence of gene expression on the androgen receptor (AR), stem cell factor (SCF), steroidogenic acute regulatory protein (StAR) and so on.To investigate the molecular mechanisms of PCOS’s follicle development and promotion regulated by Dianji Recipe in ovarian level, and then set up an experimental foundation for clinical application and dissemination.Methods:Step1. Experimental group:to establish a kind of PCOS rats model---take a21-day-old female SD rats and give subcutaneous injection of6mg/100g DHEA and0.2mL of injectable Soybean oil which begins at23-day-old stage and lasts for30days.Matched group: give subcutaneous injection of0.2mL of injectable Soybean oil for a21-day-old female SD rats,begin at23-day-old stage and last for30days.Step2. Take6-week-old female SD rats and use intragastric administration respectively by twice one day and every12hours. In the fourth day morning, Irrigate all-day dose once and obtain drug-containing sera through centrifugation1hour later for standby.Step3. Kill the PCOS rats for both group,obtain graafian follicle from ovaries and then release granulosa cells. After72hours cell culture in a bottle,take a record for two groups about the level of the E2/P and the influence caused by AR, StAR、SCFmRNA.Step4. join the drug-containing serum into cell culture mediums respectively,take a closer observation for below items:1)effect of Dianji Recipe on the secretion of granulosa cells E2/P)effect of Dianji Recipe on the AR、StAR、SCFmRNA.Meanwhile,analyses the relationship between Dianji Recipe and secretion of granulosa cell,study the influence of o Dianji Recipe n the molecules inside the ovaries of PCOS ratsResults:1. Comparing with the matched group,the weight of PCOS rat in experimental group is lost, organ coefficient of ovaries is increased, Uterine organ coefficient is decreased.Also the surface of the ovary is pale with thicker envelop,larger volume and polycystic ovary. The follicle with cystic dilatation is raised and rare corpus luteum,the levels of serum T, A, E2and P are raised. 2. PCOS rat in experimental group-the secretion of ovarian granular cells--E2is decreased and P is increased, ARmRNA is increased and SCF、StARmRNA is decreased.3. Dianji Recipe can stimulate the secretion for E2and reduce the of the secretion of P of ovarian granular cells--E2.It can also decrease the mRNA expression of AR and increase the mRNA of StAR&SCF.According to above experimental results, Dianji Recipe can improve the secretion of granulosa cells by affecting AR,StAR and SCFmRNA.Conclusion:1.successfully establish pathological PCOS rats model by the induction of DHEA and study from the filed of the oestrous cycle, ovarian morphology, pathology,endocrinology and so on. Experiment shows very similar with PCOS.2. Successfully establish vitro culture system of PCOS rat granulosa cells and study the system from various filed,such as cell growth, secretion, related gene expression. Experiment shows that in vitro conditions, can still maintain the features of PCOS.3. Experiment shows that under the control of reproductive axis, by the influence of Dianji Recipe on decreasing ARmRNA and increasing the StAR&SCF mRNA,the secretion function of granulosa cells can be gradually returned to normal.By improve abnormal condition of local ovarian autocrine and paracrine factors,ultimately achieve the goal of regulation of ovarian function.