摘要
目的观察葆青颗粒对卵巢储备功能下降(diminished ovarian reserve,DOR)模型大鼠卵泡发育的影响,探讨葆青颗粒通过PI3K/AKT/mTOR信号通路治疗DOR的作用机制。方法纳入动情周期正常的雌性SD大鼠60只,使用随机数字表法选取10只作为正常组,其余50只大鼠采用一次性腹腔注射环磷酰胺75 mg/kg诱导DOR模型。按随机数字表法将造模成功的50只大鼠分为模型组(等剂量生理盐水,灌胃)、葆青颗粒低剂量组(3 g/kg,1次/d,灌胃)、葆青颗粒中剂量组(6 g/kg,1次/d,灌胃)、葆青颗粒高剂量组(9 g/kg,1次/d,灌胃)及补佳乐组(0.21 mg/kg,1次/d,灌胃),每组各10只,连续灌胃21 d。HE染色观察卵巢及子宫形态学变化,ELISA检测血清雌二醇(estradiol,E_(2))、卵泡刺激素(follicle stimulating hormone,FSH)、抗苗勒管激素(anti-Mullerian hormone,AMH)水平;Western Blot及免疫组化法检测大鼠卵巢组织磷脂酰肌醇3-激酶(phosphoinositide 3-kinase,PI3K)、蛋白激酶B(protein kinase B,AKT)、哺乳动物雷帕霉素靶蛋白(Mammalian Target of Rapamycin,mTOR)蛋白表达。结果与空白组比较,模型组大鼠动情周期紊乱、卵巢湿重下降、闭锁卵泡增加,血清FSH浓度升高,E_(2)及AMH浓度降低(P<0.05),卵巢组织PI3K、AKT、mTOR蛋白表达降低(P<0.05)。与模型组比较,葆青颗粒各剂量组动情周期恢复、卵巢湿重增加(P<0.05),且葆青颗粒中、高剂量组及补佳乐组血清FSH浓度下降、E_(2)、AMH浓度升高(P<0.05),卵巢组织PI3K、AKT、mTOR蛋白表达升高(P<0.05)。结论葆青颗粒可能通过激活PI3K/AKT/mTOR信号通路改善DOR大鼠卵巢功能。
Objective To observe the effect of Baoqing granules on follicular development in rats with diminished ovarian reserve(DOR),and to explore the mechanism of Baoqing granules in treating DOR through the PI3K/AKT/mTOR signaling pathway.Methods Sixty female SD rats with normal estrous cycles were included,and 10 rats were selected as the normal group using a random number table method.The remaining 50 rats were induced with DOR model by one-time intraperitoneal injection of 75 mg/kg cyclophosphamide.50 successfully modeled rats were divided into a model group(equal dose of physiological saline,gavage),a low dose group of Baoqing granules(3 g/kg,once a day,gavage),a medium dose group of Baoqing granules(6 g/kg,once a day,gavage),a high dose group of Baoqing granules(9 g/kg,once a day,gavage),and a Bujiale group(0.21 mg/kg,once a day,gavage)using a random number table method.Each group had 10 rats,and they were continuously gavaged for 21 d.HE staining was used to observe the morphological changes of the ovaries and uterus,ELISA was used to detect serum estradiol(E_(2)),follicle stimulating hormone(FSH),anti-Mullerian hormone(AMH)levels,Western Blot and immunohistochemistry were used to detect phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT),mammalian target of rapamycin(mTOR)protein expression.Results Compared with the blank group,the model group rats showed estrous cycle disorder,decreased ovarian wet weight,increased atresia follicles,increased serum FSH concentration,decreased E_(2) and AMH concentration(P<0.05),and decreased expression of PI3K,AKT,and mTOR protein in ovarian tissue(P<0.05).Compared with the model group,the estrous cycle of each dose group of Baoqing Granule recovered and the ovarian wet weight increased(P<0.05),while the serum FSH concentration decreased,E2 and AMH concentrations increased(P<0.05)in the middle,high dose group of Baoqing granules and Bujiale group.and the expression of PI3K,AKT,and mTOR proteins in ovarian tissue increased(P<0.05).Conclusion Baoqing granules may improve ovarian function in DOR rats by activating the PI3K/AKT/mTOR signaling pathway.
作者
刘燕
周晓宇
汪明皓
王永周
LIU Yan(Department of Gynaecology,the Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University,Luzhou 646000,China)
出处
《牡丹江医学院学报》
2023年第6期14-20,共7页
Journal of Mudanjiang Medical University
基金
泸州市科技局应用基础项目(2022-JYJ-119)
四川省中医药管理局项目(2021MS105)。