基于线粒体能量代谢途径探讨资癸益冲方改善早发性卵巢功能不全小鼠卵巢功能的作用机制  

Study on the mechanism of Zigui Yichong Formula in improving ovarian function of POI mice based on mitochondrial energy metabolism pathway

作　　者：刘雪平 宿滋 李荣霞 唐思玲 段彦苍[1,2,3] 马惠荣 LIU Xue-ping;XIU Zi;LI Rong-xia;TANG Si-ling;DUAN Yan-cangg;MA Hui-rong(Hebei University of Chinese Medicine,Shijiazhuang 050091,China;Hebei Collaborative Innovation Center of Integrated Chinese and Western Medicine on Reproductive Disease,Shijiazhuang 050091,China;Hebei Key Laboratory of Integrative Medicine on Liver-kidney Patterns,Shijiazhuang 050091,China)

机构地区：[1]河北中医学院,石家庄050091 [2]河北省中西医结合生殖疾病协同创新中心,石家庄050091 [3]河北省中西医结合肝肾病证研究重点实验室,石家庄050091

出　　处：《中华中医药杂志》2023年第5期2361-2366,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：河北省自然科学基金项目(No.2022423334);河北省在读研究生创新能力培养资助项目(No.CXZZBS2023141);河北中医学院优秀创新团队建设项目(No.YXTD2022003)。

摘　　要：目的:探讨资癸益冲方通过线粒体能量代谢途径改善早发性卵巢功能不全(POI)模型小鼠卵巢功能的作用与机制。方法:80只SPF级雌性C57BL/6N小鼠随机分为正常组、模型组、西药(辅酶Q10)组和中药(资癸益冲方)组,每组20只。除正常组外,腹腔注射环磷酰胺(CTX)建立POI模型小鼠,造模成功后,各组灌胃给予相应药物。HE染色观察卵巢组织形态学,放射免疫法检测血清雌二醇(E_2)、卵泡刺激素(FSH)和黄体生成素(LH)水平,透射电镜观察卵巢颗粒细胞线粒体结构,比色法测定颗粒细胞三磷酸腺苷(ATP)含量,免疫组化法和Western Blot法检测与线粒体能量相关的过氧化物酶体增殖物激活受体γ共激活因子1α(PGC-1α)、核呼吸因子1(NRF1)和线粒体转录因子A(TFAM)蛋白表达水平。结果:与正常组比较,模型组小鼠卵巢各级卵泡颗粒细胞层数减少,闭锁卵泡增多,E_2水平下降(P<0.05),FSH、LH水平升高(P<0.01),颗粒细胞线粒体数量减少,出现嵴断裂及空泡化改变,外膜不完整,ATP含量下降(P<0.05),PGC-1α、NRF1、TFAM蛋白表达下降(P<0.01);与模型组比较,中药组小鼠卵巢各级卵泡颗粒细胞层数增多,闭锁卵泡减少,E_2水平升高(P<0.01),FSH、LH水平下降(P<0.01),颗粒细胞线粒体结构改善,ATP含量升高(P<0.05),PGC-1α、NRF1、TFAM蛋白表达升高(P<0.05,P<0.01)。结论:资癸益冲方可以改善线粒体能量代谢减少卵巢颗粒细胞的凋亡保护POI模型小鼠卵巢功能,其机制可能与激活PGC-1α/NRF1/TFAM信号通路有关。Objective:To investigate the efets and mechanism of Zigui Yichong Formula(ZGYCF)on improving ovarian function of premature ovarian insufficiency(POD)model mice through mitochondrial energy metabolism pathway.Methods:Eighty SPF female C57BL J6N mice were randomly divided into normal group,model group,westerm medicine(coenzyme Q10)group and traditional Chinese medicine(ZGYCF)group,with 20 mice in each group.Except the normal group,POI model mice were established by intraperitoneal injection of cyclophosphamide(CTX).After successful modeling,mice in the each group were gavaged with corresponding drug.Ovarian histomorphology appearance stained by hematoxylin eosin(HE).The serum levels of E,FSH and LH were detected by radioimmunoassay.The mitochondrial structure of ovarian granulosa cells was observed by transmission electron microscopy.The content of adenosine triphosphate(ATP)in granulosa cells was detected by colorimetry.The protein expressions of PGC-1α,NRFI and TFAM related to mitochondrial energy were detected by immunohistochemistry and Western Blot.Results:Compared with the normal group,in the model group,the number of granulosa cell layers at all levels of ovarian fllicles decreased,the number of atretic flliles increased.The level of E_(2) decreased(P<0.05),the levels of FSH and LH increased(P<0.01).The number of mitochondria in granulosa cells decreased,the changes of crista fracture and vacuolization occurred,the outer membrane was incomplete,and the content of ATP decreased(P<0.05).The protein expressions of PGC-1α,NRF1 and TFAM were decreased(P<0.01).Compared with the model group,in the traditional Chinese medicine group,the number of granulosa cell layers increased,the atretic ollicle decreased.The E,level increased(P<0.01),the levels of FSH and LH decreased(P<0.01).The mitochondrial structure of granulosa cells improved,and the content of ATP increased(P<0.05).The protein expressions of PGC-1 x,NRF1 and TFAM were increased(P<0.05,P<0.01).Conclusion:ZGYCF can reduce the apoptosis of ovarian granulosa

关 键 词：早发性卵巢功能不全 资癸益冲方 颗粒细胞 线粒体 能量代谢 

分 类 号：R285.5[医药卫生—中药学]