益经汤对环磷酰胺所致卵巢早衰小鼠的治疗作用研究

Study on the therapeutic effect of compound Yijing decoction on cyclophosphamide-induced premature ovarian failure in mice

目的 探究中药复方益经汤对环磷酰胺(CTX)所致卵巢早衰(POF)小鼠的治疗作用。方法 腹腔注射CTX建立POF小鼠模型,随机分为模型组、阳性对照组、益经汤低、高剂量组和空白组,每组6只,灌胃给药30 d。检测小鼠动情周期、体质量,卵巢、子宫指数和组织形态变化;ELISA检测血清中雌二醇(E_2)、促卵泡刺激素(FSH)、促黄体生成素(LH)、抗缪勒氏管激素(AMH)、孕酮(PROG)的浓度;免疫组织化学法(IHC)检测卵巢PI3K、AKT、FOXO3A蛋白表达水平。结果 益经汤干预模型小鼠后,动情周期趋向正常,高剂量组体质量、卵巢指数、子宫指数皆显著升高(P<0.05);卵巢体积增大,初级和次级卵泡数量增多,细胞纤维化改善;子宫内膜增厚,恢复子宫腺上皮细胞排列;阳性对照组和益经汤高剂量组血清E_2、AMH、PROG均升高(P<0.05,P<0.01),FSH、LH显著下降(P<0.01)。益经汤组和阳性对照组PI3K、AKT、FOXO3A蛋白表达皆显著降低(P<0.01)。结论 益经汤可有效改善动情周期,恢复卵巢和子宫的功能,可提高血清E_2、AMH、PROG和降低FSH、LH的表达,机制可能与下调PI3K、AKT和FOXO3A蛋白表达有关。

Objective To explore the therapeutic effect of traditional Chinese medicine compound Yijing decoction on cyclophosphamide(CTX)-induced premature ovarian failure(POF)in mice.Methods The POF mouse model was established by intraperitoneal injection of CTX,and was randomly divided into the model group,positive control group,low and high doses of Yijing decoction groups,and blank group,with 6 mice in each group.The drug was administered by gavage for 30 d.The estrous cycle,body weight,ovarian and uterine indices,and changes in ovarian and uterine tissues of mice were measured.The concentration of serum E2,FSH,LH,AMH and PROG were detected with ELISA.The levels of ovarian PI3K,AKT,and FOXO3A protein were detected by immunohistochemistry(IHC).Results After intervention with Yijing decoction in mice with POF,the estrus cycle tended to be normal.The body weight,ovarian index,and uterine index significantly increased in the high-dose group mice(P<0.05).The ovarian volume and the number of primary and secondary follicles were increased,and the phenomenon of interstitial and fibrosis was improved.The endometrium was significantly thickened,and the arrangement of endometrial glandular epithelial cells was restored.The levels of serum E2,AMH and PROG were increased in both the positive control group and the high-dose Yijing decoction group(P<0.05,P<0.01),while levels of FSH and LH were significantly decreased(P<0.01).Meantime,the expression of PI3K,AKT and FOXO3A proteins was significantly reduced in both two groups(P<0.01).Conclusion Yijing decoction can effectively improve the motility cycle and restore the function of ovary and uterus,increase the levels of serum E2,AMH,PROG,and decrease the expression of FSH and LH,which may be related to downregulation of the expression of PI3K,AKT and FOXO3A proteins.