益肾调经方对卵巢储备功能减退大鼠卵巢组织PTEN/PI3K/AKT信号通路的影响

Effect of Yishen Tiaojing Prescription on PTEN/PI3K/AKT Signaling Pathway in Ovarian Tissue of Rats with Decreased Ovarian Reserve

目的:探讨益肾调经方对卵巢储备功能减退大鼠卵巢组织PTEN/PI3K/AKT信号通路的影响,明确其改善卵巢功能的作用机制。方法:雌性SD大鼠随机分为模型组、激素序贯(补佳乐0.1 mg/kg+黄体酮胶囊21 mg/kg)组及益肾调经方高(9.2 g/kg)、中(4.6 g/kg)、低(2.3 g/kg)剂量组,连续14 d灌服雷公藤多苷片混悬液50 mg/(kg·d)造模,造模结束后连续给予相应药物治疗15 d。另设正常对照组。阴道脱落细胞学检测确认其处于动情前期并处死大鼠;计算各组大鼠性腺指数;HE染色观察卵巢一般情况及各级卵泡的变化;免疫组化染色及Western Blot检测PTEN、PI3K、AKT、p-AKT蛋白表达;qRT-PCR检测PTEN、PI3K、AKT mRNA表达。结果:与正常对照组比较,模型组动情周期紊乱,性腺指数显著降低,卵巢各级生长卵泡明显减少,PTEN mRNA及蛋白表达显著上调,PI3K、AKT、p-AKT mRNA和(或)蛋白表达显著下调;与模型组比较,益肾调经方组动情周期显著改善,大鼠性腺指数显著升高,卵巢各级生长卵泡增多,PTEN mRNA及蛋白表达显著下调,PI3K、AKT、p-AKT mRNA和(或)蛋白表达显著上调。结论:益肾调经方能有效改善DOR大鼠的卵巢功能,其作用机制可能与PTEN/PI3K/AKT通路有关。

Objective:To explore the effect of Yishen tiaojing prescription on PTEN/PI3K/AKT signaling pathway in ovarian tissue of rats with decreased ovarian reserve function,and to clarify its mechanism of improving ovarian function.Methods:60 female SD rats were randomly divided into model group,hormone sequence(Bujiale 0.1 mg/kg and progesterone capsule 21 mg/kg)group and high(9.2 g/kg),middle(4.6 g/kg),low(2.3 g/kg)dose groups of Yishen tiaojing prescription.The rats were treated with tripterysium polyglycoside tablet suspension 50 mg/(kg·d)for 14 days,and then treated with corresponding drugs for 15 days.Normal control group was also set.Vaginal exfoliative cytology confirmed that it was in proestrus and the rats were killed,the gonadal index of rats in each group was calculated,the general condition of ovary and the changes of follicles at all levels were observed by HE staining,the proteins expressions of PTEN,PI3K,AKT and p-AKT were detected by immunohistochemical staining and Western Blot,the mRNA expression levels of PTEN,PI3K and AKT were detected by qRT-PCR.Results:Compared with the normal control group,the estrous cycle of the model group was disordered,the gonadal index decreased significantly,the growth follicles at all levels of the ovary decreased significantly,the expression of PTEN mRNA and protein was significantly up-regulated,and the expressions of PI3K,AKT,p-AKT mRNA and/or proteins was significantly down-regulated.Compared with the model group,the estrous cycle of Yishen tiaojing prescription group was significantly improved,the gonadal index significantly increased,the number of growing follicles at all levels of ovary increased,the expressions of PTEN mRNA and protein significantly down-regulated,and the expression of PI3K,AKT,p-AKT mRNA and/or protein significantly up-regulated.Conclusion:Yishen tiaojing prescription can effectively improve the ovarian function of DOR rats,and its mechanism may be related to PTEN/PI3K/AKT pathway.