温中补肾方对多囊卵巢综合征改善作用及对雌性生殖干细胞增殖的影响  

Effects of Wenzhong Bushen Formula on PCOS and proliferation of female reproductive stem cells

作　　者：王春红 于普光 孙亚群 彭庆杰 李雪 吴昕 马会明[1] 李广永[3] 杜小利[2] 何瑞 WANG Chun-hong;YU Pu-guang;SUN Ya-qun;PENG Qing-jie;LI Xue;WU Xin;MA Hui-ming;LI Guang-yong;DU Xiao-li;HE Rui(Key Laboratory of Fertility Maintenance,Ministry of Education,Basic Medical College,Ningxia Medical University,Yinchuan 750004,China;Key Laboratory of Hui Medicine Modernization,College of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750004,China;General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院,生育力保持教育部重点实验室,银川750004 [2]宁夏医科大学中医学院,回医药现代化教育部重点实验室,银川750004 [3]宁夏医科大学总医院,银川750004

出　　处：《中华中医药杂志》2022年第6期3564-3568,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：宁夏重点研发计划项目(No.2020BFG02010);国家自然科学基金项目(No.31860290);宁夏科技创新领军人才培养项目(No.2020GKLRLX11);宁夏医科大学校级科研项目(No.XT2019017)。

摘　　要：目的:探究温中补肾方对多囊卵巢综合征(PCOS)的改善作用及机制。方法:4~6周龄C57BL/6J雌性小鼠,连续灌胃来曲唑联合高脂饲料喂养,建立PCOS模型。成模后给予不同剂量的温中补肾方,期间测体质量及糖耐量试验(OGTT)等。取材后ELISA检测睾酮(T)等激素水平;HE染色检测卵巢形态改变;免疫组化法检测MVH、AMH蛋白表达;免疫荧光法共染雌性生殖干细胞(FGSCs)标志物;Western Blot法检测卵巢组织中MVH、PCNA、p-mTOR、p-AMPK等蛋白。结果:与PCOS模型组比较,温中补肾方干预后,PCOS小鼠体质量、OGTT及T等水平显著改善(P<0.05,P<0.01),卵巢多囊病变减轻,具有增殖性的FGSCs数目显著增多(P<0.05),p-mTOR、p-AMPK显著上调(P<0.01,P<0.05)。结论:温中补肾方不仅可以改善PCOS糖脂代谢异常及卵巢多囊病变,还可以激活FGSCs增殖并激活其功能,其作用可能与AMPK、mTOR磷酸化有关。Objective:To explore the improvement effect and mechanism of Wenzhong Bushen Formula on polycystic ovarian syndrome(PCOS).Methods:Female C57BL/6J mice aged 4~6 weeks were fed with letrozole and high-fat diet to establish PCOS model.After successful modeling,different doses of Wenzhong Bushen Formula were given,body weight and oral glucose tolerance test(OGTT)were detected.The levels of testosterone(T)and other hormones were detected by ELISA.The morphological changes of ovary were detected by HE staining;The protein expressions of MVH and AMH were detected by immunohistochemistry;The markers of female germline stem cells(FGSCs)were co stained by immunofluorescence;MVH,PCNA,p-mTOR and p-AMPK were detected by Western Blot.Results:After the intervention,compared with PCOS model group,the weight,OGTT,T levels improved(P<0.05,P<0.01),ovarian polycystic lesions decreased.In addition,the number of proliferative FGSCs increased significantly(P<0.05),further study showed that p-mTOR and p-AMPK were up-regulated(P<0.01,P<0.05).Conclusion:Wenzhong Bushen Formula can not only improve PCOS glucose and lipid metabolism and polycystic ovary disease,but also activate the proliferation and function of FGSCs.Its effect is possibly related to the phosphorylation of AMPK and mTOR and the promotion of FGSCs proliferation.

关 键 词：多囊卵巢综合征 温中补肾方 雌性生殖干细胞 AMPK MTOR 磷酸化 机制 动物模型 

分 类 号：R285.5[医药卫生—中药学]