| Objective:The objective of this thesis is to observe the effects of Huluantang on the follicular development and the quality of occytes in rats under GnRHa controlled ovarian hyperstimulation, to explore the related mechanism in both cellular and molecular levels, and to provide scientific basis for effectiveness of Huluantang by assisted reproductive technology.Methods:After receiving GnRHa pituitary down regulation pretreatment which imitates the human long period of COH, a number of rats were separated into a so-called model group treated with HMG/HCG superovulation drugs and so-called Chinese medicine groups treated with HMG/HCG superovulation drugs and different doses of Chinese medicine of Huluantang. The natural cycle of the rats were compared to a normal group receiving no medicine. The following indicators were observed on the day of HCG injection:morphology of ovary by means of light microscope and electron microscope; cell apoptosis of ovary through TUNEL; contents of FSH, LH, E2, P in serum by radioimmunoassay; contents of cytokines in ovary by antibody array; expression of FSHR and LHR in ovary through immunohistochemistry.Results:1. Compared with those in the normal group:the increase of the ovary index in the model group had statistical significance (P<0.05); the decrease of the rate of high quality follicular had statistical significance obviously (P<0.01); the ultrastructure of oocyte was abnormal; the increase of cell apoptosis of ovary had statistical significance obviously (P<0.01); the levels of ACTH, BDNF, basic-FGF, CSK, GM-CSF, GH, GHR,IL-1β, IL-12/IL-23p40, EG-VEGF/PK1,TIMP-1, TIMP-2, TIMP-3, NGF-β, PDGF-AA in ovary were decreased above1.5times, the levels of Leptin (OB), IFN-y, Fas/TNFRSF6, TRAIL in ovary were increased above1.5times; the decrease of the expression of FSHR in ovary had statistical significance (P<0.05).2. Compared with those in the model group, the increase of the ovary index in the middle dose group had statistical significance obviously (P<0.01); the increase of the ovary index in the low dose and high dose groups had no statistical significance (P>0.05); the decrease of cell apoptosis of ovary in the middle dose group had statistical significance (P <0.05); the decrease of cell apoptosis of ovary in the low dose and high dose groups had statistical significance obviously (P<0.01); the increase of the rate of high quality follicular in the middle close and high dose groups had statistical significance (P<0.05); the increase of the rate of high quality follicular in the low dose group had no statistical significance (P>0.05); the increase of the content of E2in serum in the middle dose group had statistical significance obviously (P<0.01); the increase of the content of E2in serum in the high dose groups had statistical significance (P<0.05); the increase of the content of E2in serum in the low dose group had no statistical significance (P>0.05); the increase of the expression of FSHR in ovary in the high dose group had statistical significance (P<0.05); the increase of the expression of LHR in ovary in the middle dose group had statistical significance obviously (P<0.01); the increase of the expression of FSHR in ovary in the high dose group had statistical significance (P<0.05); the levels of ACTH, BDNF, basic-FGF, CSK, EG-VEGF/PK1, GFR-α1, GM-CSF, GHR, IL-1β, IL-12/IL-23p40, NGF-β, PDGF-AA, TGF-β1, TGF-β33, TIMP-3in ovary increased above1.5times in the middle dose group; the levels of Fas/TNFRSF6, IFN-γ, Leptin (OB), TNF-a in ovary were decreased above1.5times in the middle dose group; the total live fetus in the middle dose group increased above1.5times.Conclusions:1. Compared with those in normal natural cycle, the cell apoptosis of ovary increased and the rate of high quality follicular and quality of occytes decreased, and the contents of cytokines in ovarian microenvironment were abnormal for rats under GnRHa superovulation. The results suggest some of the adverse effects of GnRHa superovulation on follicular development.2. The Chinese medicine of Huluantang can increase the expression of some cytokines in ovarian microenvironment which can promote follicular development and the quality of occytes, such as IL-1β,GHR, TGF-β1, and decrease the expression of some cytokines such as IFN-γ, TNF-α and Leptin. The results suggest that Huluantang can improve the ovarian microenvironment in rats under GnRHa superovulation.3. The Chinese medicine of Huluantang can decrease the ovarian cell apoptosis in rats under GnRHa superovulation. The reason may be the decrease of the expression of IFN-y and TNF-a in the ovarian microenvironment.4. The Chinese medicine of Huluantang can increase the content of E2in serum, which suggests Huluantang can improve the endocrine environment in rats under GnRHa superovulation. The reason may be the improvement of the cellular and molecular environment in ovary by Huluantang.5. The Chinese medicine of Huluantang can increase the expression of FSHR and LHR in ovary, which may improve the ovarian response in rats under GnRHa superovulation. The reasons may be the improvement of ovarian microenvironment and endocrine environment.6. Through the above mechanism, the Chinese medicine of Huluantang can promote the follicular development, the weight of ovary and the quality of occytes in rats under GnRHa superovulation, through which more high quality occytes and live fetus can be obtained. The results suggest that Huluantang may increase the success rate of assisted reproductive technology (ART).
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