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目的:研究补肾调冲方对半乳糖致POF大鼠卵巢功能的调节,探讨补肾调冲方通过调控卵巢始基卵泡募集因子GDF-9及AMH调节卵巢功能及治疗POF的机制。方法:建立半乳糖致POF大鼠模型,将造模成功的POF大鼠随机分为模型组与治疗组,每组各20只,另设正常对照组。于PND80开始,治疗组大鼠每日灌服补肾调冲方水溶液,模型组及空白组大鼠每日灌服等体积生理盐水,共4周。实验结束后光镜下计数各组大鼠卵巢内各级卵泡数目,ELISA法检测各组大鼠血清FSH、LH及E2水平,Real-time PCR法检测各组大鼠卵巢内GDF-9及AMH mRNA表达水平。结果:①补肾调冲方可使POF大鼠动情周期趋于正常,并能明显降低血清FSH、LH水平(P<0.05),提高E2水平(P<0.05),从而改善POF大鼠血清高促性腺激素低雌激素状态;组织形态学检测结果显示治疗组大鼠卵巢体积较POF模型组增大,各级卵泡生长较好,颗粒细胞层次多,黄体数目明显增多;始基卵泡、窦前卵泡及窦状卵泡数目均较模型组增多,闭锁卵泡数目则明显减少。②与正常组比较,POF大鼠卵巢GDF-9及AMH mRNA水平均明显降低(P<0.01),而治疗组大鼠卵巢GDF-9及AMH mRNA水平较模型组明显升高(P<0.05或P<0.01)。结论:补肾调冲方治疗POF的作用机制之一可能是通过上调卵巢局部始基卵泡募集因子GDF-9及AMH的表达,促进卵泡的生长发育,抑制卵泡的过度耗竭与闭锁,恢复始基卵泡的正常募集状态,从而抑制卵巢功能的过早衰竭。
OBJECTIVE: To investigate the effect of Bushen Tiaochong Recipe on the ovarian function of POF-induced rats induced by galactose, and to explore the mechanism of Bushen Tiaochong Recipe in regulating ovarian function and treating POF by regulating the recruitment of ovarian primordial follicles GDF-9 and AMH. Methods: POF rat model induced by galactose was established. POF rats with successful modeling were randomly divided into model group and treatment group, with 20 rats in each group and another normal control group. At the beginning of PND80, the rats in the treatment group were fed with Bushen Tiampan Fang aqueous solution daily. The rats in the model group and the blank group were given an equal volume of normal saline daily for 4 weeks. At the end of the experiment, the number of follicles in each ovary of each group was counted under light microscope. Serum FSH, LH and E2 levels were measured by ELISA. The levels of GDF-9 and AMH in ovaries of rats in each group were detected by Real-time PCR mRNA expression level. Results: Bushen Tiaochong recipe can make the oestrous cycle of POF rats normal and reduce serum FSH and LH levels (P <0.05) and E2 levels (P <0.05) Hormone low estrogen status; histological examination showed that the treatment group rats ovarian volume than the POF model group increased follicle growth at all levels better, more granular cell layers, the number of corpus luteum significantly increased; primordial follicles, preantral follicles and The number of antral follicles were more than the model group, the number of atresia follicles was significantly reduced. ②Compared with the normal group, the levels of GDF-9 and AMH mRNA in ovary of POF rats were significantly decreased (P <0.01), while the levels of GDF-9 and AMH mRNA in ovary of POF rats were significantly higher than those in model group (P <0.05 or P <0.01). Conclusion: One of the mechanisms of Bushen Tiaochong Recipe in treating POF may be through the up-regulation of the expression of GDF-9 and AMH in ovary, promoting the growth and development of follicles, inhibiting the excessive depletion and atresia of follicles, restoring the primordial follicles Normal recruitment state, thereby inhibiting premature ovarian failure.