| Background and ObjectivePremature ovarian failure has become one of the diseases which seriousely endanger the health of women, the age of onset is more and more younger,the incidence is increasing. Although with the rapid development of science and technology,we have made some progress on the diagnosis and treatment of premature ovarian failure,but because of its complex etiology and pathogenesis,it is difficult to make a early diagnosis.POF can cause extensive complications.We have not found the ideal treatment.Therefore,it is very important and necessary to prove its causes further and look for ways to effectively intervene in the occurrence and development of premature ovarian failure.There is currently no ideal treatment for POF.The modern medical mainly uses of hormone replacement therapy,induced ovulation, immunosuppressive therapy,self or allogeneic ovarian transplantation and other methods. In recent years, isoflavones have been shown to have a better theraepeutic effect of premature ovarian failure.Pomiferin is a kind of isoflavone which is extracted from Osage orange.lt has a week estrogenic activity. Its molecular formula is C25H24O6 and its molecular weight is 420. A lot of studies have demonstrated that pomiferin has extensive effects. It can been a powerful antioxidant, a phytoestrogen, a metal chelating agent,a antimicrobial agent.It also plays the role of cell cycle regulation.There are lots of domestic and international researches which show that:pomiferin is a phytoestrogen which can mimic the effect of endogenous estrogens.Many estrogens usually bind to ERa which can act as a strong stimulus for breast cell proliferation that may result in cancer.Compared with them, pomiferin binds to ERβwith higher affinity. ERβcan oppose the effect of ERa. So pomiferin can reduce the proliferation of breast, the incidence of uterine fibroids, blood clots and other adverse reactions.The specific effect for pomiferin to treat premature ovarian failure is still not clear, especially the mechanism on pomiferin how to intervent POF is still scarce.Modern medicine generally considered that etiology of POF is very complex and the exact cause is not yet clear. There are a lot of causes, such as genetic factors, immune factors, radiation, virus infection and so on. One of the causes is the reduction of primordial follicle reserves, accelerated apoptosis, follicular atresia. The study on POF is continuning. In recent years, apoptosis is beginning to attract attention.Apoptosis is a initiative progress in which the cells begin to aging and death.This progress is affecte'd by gene regulation according to certain procedures. Apoptosis provides a new basis for the etiology of POF.In the ovary, the procreation and dividation of follicle cells are regulated by apoptosis. It can be said that apoptosis accelerates follicular atresia directly. The main way to reduce a mature T cell activation and apoptosis is the combination of Fas and Fas-L. Bcl-2 plays a contrast role with Fas. It is said that Bcl-2 can inhibit Fas-mediated apoptosis.Therefore, based on the above analysis, this study decides to evaluate the Intervention effect of pomiferin on POF.MethodThe animal model of autoimmune POF was established, we speculate the regulation of follicle apoptosis of pomiferin on autoimmune POF.1.Animal model construction, group classifiaction and administration.We injectde a mouse zone pellucida with the 330-342 amino acid sequence as immunogen to induce autoimmune POF. The mice were randomly divided into six groups including control group,adjuvant group,model group,promiferin low-dosage group,middle-dosage group and high-dosage group. 2.The ovarian morphology and estrous cycle of mice in each group were observed, electronic balance are used to weigh the body weight,ovarian weight, thymus weight and spleen weight after treatment and to compare their weight exponent.3. Enzyme-linked immunosorbent assay were used to measure the peripheral blood FSH, E2 levels.4. RT-PCR method was used to determine the Fas-L and Bcl-2mRNA expression of the ovaian tissue.5. Immunohistochemistry was used to measure the Fas-L and Bcl-2 protein expression in the ovarian tissue.6. Western -blot was used to measure the Fas-L and Bcl-2 protein expression in the ovarian tissue.Results'Model group:1. Estrous cycle:the estrous cycle became disorder slowly, the diestrus was prolonged, the vaginal smear mainly showed a large number of white blood cells and a small amount of epithelial cells.2. Ovarian morphology:less mature follicles and infiltration of many lymphocytes and plasma cells were observed in the ovarian stroma.3. The weight exponents of body, ovary, thymus and spleen have fallen.4. The level of E2 in peripheral blood decreased and FSH significantly increased.5. The expression of Fas-L increased and expression of Bcl-2 reduced significantly. Pomiferin group:1. Estrous cycle: the estrous cycle became mild disorder, the diestrus was slight prolonged.It shows mainly proestrus. The vaginal smear mainly showed a number of nuclear epithelial cells and a small number of keratinocytes.2. Ovarian morphology: there are more development and maturation of ovarian follicles. Also there are less infiltration of lymphocytes and plasma cells were observed in the ovarian stroma.3. The weight exponents of ovary and thymus were increased significantly while a slight increase in the weight exponent of body.4. The level of E2 in peripheral blood increased and FSH level significantly decreased.5. The expression of Fas-L reduced and expression of Bcl-2 increased significantly.Conclusions1. Through the testing of the above model group, the model of premature ovarian failure was successfully constructed.2. Pomiferin plays a regulatory role on the mice with autoimmune POF. It can increase the weight index of thymus.It can increase the level of E2 and decrease the level of FSH.3. Pomiferin can improve the POF caused by autoimmune response. It can increase ovarian mass index significantly; also it can make an increasfe of mature ovarian follicles.4. Pomiferin can inhibit Fas-mediated apoptosis of follicle cells, pomiferin can decrease fas-1 expression and increase Bcl-2 expression. |
PDF Full Text Request