摘要
目的探讨金丝桃苷(Hyperin)对雷公藤苷(TG)诱导的早发性卵巢功能不全(POI)小鼠卵巢储备的改善作用及机制。方法以成年雌性BALB/c小鼠为研究对象,随机分为对照组、POI模型组和Hyperin治疗组,每组40只。雷公藤苷40 mg/kg,每日2次,灌胃2周,构建POI小鼠模型,Hyperin治疗组于造模后给予Hyperin 75 mg/(kg·d),灌胃4周。称取小鼠体质量,计算性腺指数。HE染色观察卵巢组织学改变,计算各级卵泡数量。ELISA法检测血清雌二醇(E2)、促卵泡生成素(FSH)、抗苗勒管激素(AMH)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的水平。RT-qPCR和Western blot分别检测卵巢颗粒细胞中核因子E2相关因子2(Nrf-2)、血红素氧化酶-1(HO-1)、Caspase3、Bcl-2、Bax的mRNA和蛋白水平,Western blot检测颗粒细胞中磷酸化磷脂酰肌醇-3-羟激酶(p-PI3K)、磷酸化蛋白激酶B(p-Akt)的蛋白水平。H2DCFDA探针法检测颗粒细胞活性氧簇(ROS)水平。TUNEL法检测颗粒细胞凋亡水平。结果与POI模型组相比,Hyperin治疗组小鼠体质量和性腺指数均上升(P<0.05);卵巢病理损伤减轻,各级卵泡数和黄体数均增加(P<0.05);血清E2、AMH、SOD、CAT水平升高,FSH水平降低(P<0.05);卵巢颗粒细胞中的Nrf-2、HO-1、p-PI3K、p-Akt、Bcl-2表达上升,Caspase3、Bax表达下降(P<0.05);ROS水平降低(P<0.05);TUNEL显示颗粒细胞凋亡降低(P<0.05)。结论Hyperin通过Nrf-2/HO-1抗氧化应激和PI3K/Akt抗凋亡途径改善TG诱导的POI小鼠卵巢储备功能的下降。
Objective To investigate the effect and mechanism of hyperin on the improvement of ovarian reserve of tripterygium glycosides(TG)-induced primary ovarian insufficiency(POI)in mice.Methods Adult female BALB/c mice were used as research subjects and were randomly assigned to the control group,POI model group and hyperin treatment group,with 40 mice in each group.TG was given at 40 mg/kg twice a day by gavage for 2 weeks to create the POI mouse model.Mice in the hyperin treatment group were given hyperin at 75 mg/(kg·d)by gavage for4 weeks after the model was established.The body mass of the mice was weighed and the gonadal index was calculated.Ovarian histological changes were observed by HE staining,and the number of follicles at all levels was calculated.Serum estradiol(E2),follicle-stimulating hormone(FSH),anti-mullerian hormone(AMH),superoxide dismutase(SOD)and catalase(CAT)were assessed with ELISA.The mRNA and protein levels of nuclear factor(erythroid-derived 2)-related factor 2(Nrf-2),heme oxygenase-1(HO-1),Caspase3,Bcl-2 and Bax in ovarian granulosa cells were measured by RTqPCR and Western blot.The protein levels of phosphorylated phosphatidylinositol 3-hydroxy kinase(p-PI3 K)and phosphorylated protein kinase B(p-Akt)were measured by Western blot.The reactive oxygen species(ROS)levels in granulosa cells were determined by H2 DCFDA.The apoptosis of granulosa cells was examined by TUNEL assay.Results Compared with mice in the POI model group,the body mass and gonadal index of hyperin-treated mice increased(all P<0.05).The pathological damage of the ovary decreased,and the number of follicles at all levels and corpora lutea increased(all P<0.05).Serum E2,AMH,SOD and CAT levels increased,and FSH level decreased(all P<0.05).At the molecular level,the expression of Nrf-2,HO-1,p-PI3 K,p-Akt and Bcl-2 in ovarian granulosa cells increased,while the expression of Caspase3 and Bax decreased(all P<0.05).ROS level decreased(P<0.05).TUNEL assay showed reduced apoptosis of granulosa cells(P<0.05).Conclusion Hyperin impr
作者
马蔚蓉
谈勇
MA Weirong;TAN Yong(Department of Gynecology,Jiangsu Province Hospital of Chinese Medicine/Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Department of Reproductive Medicine,Jiangsu Province Hospital of Chinese Medicine/Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China)
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2021年第3期458-466,共9页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(No.81873333)
江苏省卫健委临床医学中心资助项目(No.YXZXB2016006)
江苏省中医院院级课题(No.Y18068)资助。
