基于PI3K/AKT/mTOR通路研究杜仲总黄酮对多囊卵巢综合征大鼠性激素水平及卵巢组织自噬的影响  

Effects of Total Flavonoids from Eucommia ulmoides Oliv. on Sex Hormone Levels and Autophagy in Ovaries of Rats with Polycystic Ovary Syndrome Based on PI3K/Akt/mTOR Pathway

作　　者：毕晓英 许珂[1] 赵艳晓[1] 郑瑞君[2] Bi Xiaoying;Xu Ke;Zhao Yanxiao;Zheng Ruijun(Reproductive Center of the Third Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450008;Department of Gynecology,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450002)

机构地区：[1]河南中医药大学第三附属医院生殖中心,郑州450008 [2]河南省中医院妇科,郑州450002

出　　处：《中药药理与临床》2022年第1期82-86,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：2018年度河南省中医药科学研究专项课题(编号:2018ZY2100)

摘　　要：目的:探讨杜仲总黄酮基于磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)通路对多囊卵巢综合征(PCOS)大鼠性激素水平及卵巢组织自噬的影响。方法:55只SD大鼠随机取10只作为正常对照组,其余建立PCOS大鼠模型,40只大鼠建模成功,随机分为模型对照组、PI3K抑制剂BKM12040 mg/kg组、BKM12040 mg/kg+杜仲总黄酮200 mg/kg组、杜仲总黄酮200 mg/kg组,各组灌胃生理盐水或相应药物,连续干预3 w。Elisa法检测血清促黄体生成素(LH)、卵泡刺激素(FSH)、抗苗勒氏管激素(AMH)水平;HE染色观察卵巢病理学变化;流式细胞术检测卵巢组织颗粒细胞自噬率;Western blot检测卵巢组织中PI3K/AKT/mTOR通路相关蛋白及微管相关蛋白1轻链3 II(LC3 II)表达。结果:与正常对照组比较,模型对照组大鼠AMH、LH、T含量显著升高,FSH含量显著降低P<0.01);大鼠卵巢组织颗粒细胞自噬率显著升高(P<0.01),p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR比值显著降低,LC3 II/LC3 I比值显著升高(P<0.01);与模型对照组比较,抑制剂组大鼠AMH、LH、T含量显著升高,FSH含量显著降低(P<0.01),卵巢组织颗粒细胞自噬率显著升高,p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR比值显著降低,LC3 II/LC3 I比值显著升高(P<0.01);杜仲总黄酮+抑制剂组、杜仲总黄酮组大鼠AMH、LH、T含量明显降低,FSH含量明显升高(P<0.05或P<0.01),卵巢组织颗粒细胞自噬率显著降低(P<0.01),p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR比值显著升高,LC3 II/LC3 I比值显著降低(P<0.01)。模型对照组卵巢表面苍白,黄体减少,出现多个囊状扩张卵泡;杜仲总黄酮+抑制剂组、杜仲总黄酮组卵巢体积减小,表面色泽较红,黄体有所增加,囊状扩张卵泡减少;抑制剂组卵巢体积更大、黄体更少,囊状扩张卵泡更多。结论:杜仲总黄酮可能通过激活PI3K/AKT/mTOR通路降低PCOS大鼠卵巢组织自噬水平,改善其性激素水平。Objective:To investigate the effects of total flavonoids of Eucommia ulmoides Oliv.on sex hormone levels and ovarian granulosa cell autophagy in rats with polycystic ovary syndrome(PCOS)based on phosphatidylinositol-3 kinase(PI3 K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway.Methods:Ten of 55 SD rats were randomly selected as the normal control group,and the rest rats were used to establish the PCOS model.Forty PCOS rats were randomly assigned into the model control group,PI3 K inhibitor(BKM120,40 mg/kg)group,BKM120(40 mg/kg)+E.ulmoides total flavonoids(200 mg/kg)group,and E.ulmoides total flavonoids(200 mg/kg)group,with 10 rats in each group.All the groups were administrated with corresponding drugs or normal saline for three weeks.The serum levels of luteinizing hormone(LH),follicle-stimulating hormone(FSH),testosterone(T),and anti-Mullerian hormone(AMH)were measured by enzyme-linked immuno sorbent assay.HE staining was carried out for observation of the pathological changes of ovary.Flow cytometry was employed to detect the autophagy rate of ovarian granulosa cells.The protein levels of PI3 K/AKT/mTOR and microtubule-associated protein 1 light chain 3 II(LC3 II)in ovarian tissues were measured by Western blotting.Results:Compared with the normal control group,the modeling increased the serum AMH,LH,and T levels and decreased the FSH level(P<0.01).Additionally,the modeling promoted the autophagy of ovarian granulosa cells(P<0.01),decreased the p-PI3 K/PI3 K,p-AKT/AKT,and p-mTOR/mTOR ratios,and increased the LC3 II/LC3 I ratio(P<0.01).Moreover,PI3 K inhibitor aggravated such changes(P<0.01).However,E.ulmoides total flavonoids+PI3 K inhibitor and E.ulmoides total flavonoids alone lowered the AMH,LH,and T levels,elevated the FSH level(P<0.05 or P<0.01),inhibited the autophagy of ovarian granulosa cells(P<0.01),increased the p-PI3 K/PI3 K,p-AKT/AKT,p-mTOR/mTOR ratios,and decreased the LC3 II/LC3 I ratio(P<0.01).The rats in the model group showed pale ovarian surface,reduced corpus luteum,and mu

关 键 词：杜仲总黄酮 多囊卵巢综合征 性激素 自噬 磷脂酰肌醇-3激酶/丝氨酸苏氨酸蛋白激酶/哺乳动物雷帕霉素靶蛋白通路 

分 类 号：R285.5[医药卫生—中药学]