Effect Of Bushenhuoxue Decoction On Chronic Stressed Premature Ovarian Failure In Mice

Objective:By observing the effect of Bushenhuoxue decoction on chronic stressed premature ovarian failure (POF) in the model mice, evaluate the preventive and therapeutic effects of Bushenhuoxue decoction on POF.Methods:The male A/J mice and female C57BL/6 mice were cross-fertilized to generate B6AF1 mice.60 female B6AF1 mice that have normal estrous cycle were used to do animal experiments,60 mice were randomly divided into normal group (20 mice), stress group (40 mice). The mice of chronic stress POF were established. After four weeks, 10 mice were selected randomly from the normal group and stress group to evaluate the mice model, vaginal exfoliated cells stained by Papanicolaou, the ovary index and uterus index calculated, ovarian and uterine tissue morphology observed by H&E staining were together used to judge if the animal model is available. The mice of stress group that have normal estrous cycle were randomly divided into model group, traditional chinese medicine (TCM)group (Bushenhuoxue decoction, solution concentration of 3.15g/ml) and progynova group, every group mice were given corresponding medicines by oral gavage. Normal group and model group mice were administrated orally the equal volume of saline. And lasted for two weeks. We drawn the blood from all mice after oral administration, and the blood were centrifuged to measure estradiol(E2) and follicle stimulating hormone levels(FSH) by Enzyme-linked immunoadsorbent assay (ELISA). We calculated the ovary index and uterus index. Ovarian and uterus tissue morphology were observed by H&E.Results:1. Ovarian and uterine indexes of mice:Compared with the normal group, the ovary index and uterine index of every group decreased, but the model group declined markedly. The ovary index and uterus index of TCM group and progynova group were higher than the model group, which were statistically significant (P<0.05). There was no obvious difference between TCM group and progynova group (P>0.05).2. The sex hormone levels of mice:The estradiol levels of TCM group and progynova group were higher than the model group, which were statistically significant (P<0.05). There was no obvious difference between TCM group and progynova group (P>0.05). FSH levels of TCM group and progynova group were lower than the model group. But there was no significant difference upon decline of FSH between TCM group and model group(P>0.05). There was significant difference upon decline of FSH between progynova group and model group(P<0.05). There was no obvious difference upon decline of FSH between TCM group and progynova group (P>0.05).3. The ovarian morphology of mice:Ovarian follicles were clearly visible in normal group mice, morphological rule, the number of corpus luteum was more, granulosa cells were alined. But ovarian follicles were scarcein in model group, the shape was irregular. The number of corpus luteum was less, granular cell was not regular. There were more interstitial glands in granular cell, which were partly replaced by fibrous tissue. The number of ovary follicles was more in TCM group and progynova group than model group. And we could see growth follicle and mature follicle, normal follicle morphology, granular cell arrangement was regular in TCM group and progynova group.4. The uterine morphology of mice:The normal endometrium account for more two-thirds of the uterine wall was observed in normal and TCM groups. Epithelial layer was mainly secretory cells. he connective tissue of epithelial layer is loose. There was a large number of secretory granule in secretory cells cytoplasm. Lamina propria had many uterine glands. Lumens were large and regular. But the endometrium of model group was thinner, accounting for one-twice to one-thirds of the uterus wall. Epithelial layer was mainly columnar cells. There was a lack of secretions in cavity of uterine glands. Connective tissues of lamina propria were widely distributed. uterus glands are irregular and less in lamina propria. Lumens were small, and even atrophy. The endometrium of progynova group was rather thicker, accounting for one-thirds to two-thirds of the uterus wall. There were more secretory cells in epithelial layer. The connective tissue of epithelial layer is loose. There were many uterine glands in epithelial layer. Lumens were regular.Conclusion:Bushenhuoxue decoction can improve reproductive endocrine function of POF mice, promote the development of follicle, increase the endometrial receptivity. Bushenhuoxue decoction have the possible effects in the treatment of cronic stressed POF.