补肾调冲方对半乳糖致POF大鼠始基卵泡募集因子GDF-9、AMH表达的影响

Effects of Bushen Tiaochong Recipe on Primordial Follicular Recruitment Factors GDF-9 and AMH expression in galactose toxicity-induced premature ovarian failure rats

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 observe the effect of Bushen Tiaochong Recipe（BSTCR） on primordial follicular recruitment factors,GDF-9 and AMH,in galactose toxicity-induced premature ovarian failure（POF） rats.Methods：The galactose toxicity-induced POF rat model was established.POF rats were divided into model group and treated group,each 20.The treated rats were orally given BSTCR solution 6.26g/kg（equivalent human dose based on body-mass index adjustment for the adult） daily,the normal rats and POF rats given physiological saline for 4 weeks.The estrous cycle of the rats were observed.Histological changes were analyzed by light microscope as well as counting the numbers of primordial follicles（POF）,growing follicles（GF）,antral follicles（AFC） and atretic follicles（ATF）.Quantification of E2,FSH and LH were examined by ELISA.Growth differentiation factor-9（GDF-9） and anti-mullerian hormone（AMH） mRNA were measured by real-time PCR.Results：①Administration of BSTCR to POF rats significantly improved galactose toxicity-induced depressant follicle development and ovulation,increased follicle atresia,destructive ovarian histomorphology and the state of hypergonadotrophic hypoestrogenism,so the ovarian function of POF rats was regained.②Galactose toxicity caused a marked decrease in the levels of GDF-9 and AMH mRNA in ovary,which led to a increasing PMF recruitment and exhaustion.Administration of BSTCR exhibited a depressant effect on galactose-induced loss of GDF-9 and AMH.Conclusions：One of treatment mechanism of BSTCR on POF maybe the depressant effect on galactose-induced loss of GDF-9 and AMH.