青蒿琥酯对多囊卵巢综合征模型小鼠的治疗作用及机制研究  

Therapeutic efficacy and mechanism of artesunate for mouse model of polycystic ovary syndrome

作　　者：王雪玲 钟佩伶 赵志鹏 陈飞 刘鑫 刘思佳 袁烈 方路 姚千弋 杨雄 刘超[2] 成嘉坤 蔡永青[7] 李小丽[2,3] 李维宏 WANG Xueling;ZHONG Peiling;ZHAO Zhipeng;CHEN Fei;LIU Xin;LIU Sijia;YUAN Lie;FANG Lu;YAO Qianyi;YANG Xiong;LIU Chao;CHENG Jiakun;CAI Yongqing;LI Xiaoli;LI Weihong(Reproductive Medicine Center,the First Affiliated Hospital of Chongqing Medical University,Chongqing Medical University,Chongqing;Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology,Chongqing Medical University,Chongqing;Faculty of Pharmacology,College of Pharmacy,Chongqing Medical University,Chongqing;Department of Pathology,Qujing First People’s Hospital,Qujing,Yunnan;Department of Pharmacy,Dazhou Integrated TCM&Western Medicine Hospital,Dazhou,Sichuan;Clinical Medical Research Center,First Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing;Department of Pharmacy,Army Medical Center of PLA/Daping Hospital of Army Medical University,Chongqing,China)

机构地区：[1]重庆医科大学附属第一医院生殖医学中心,重庆 [2]重庆医科大学重庆高校生物化学与分子药理学重点实验室,重庆 [3]重庆医科大学药学院药理系,重庆 [4]曲靖市第一人民医院病理科,云南曲靖 [5]达州市中西医结合医院药学科,四川达州 [6]陆军军医大学(第三军医大学)第一附属医院临床研究中心,重庆 [7]陆军特色医学中心(第三军医大学大坪医院)药剂科,重庆

出　　处：《陆军军医大学学报》2025年第3期193-204,共12页Journal of Army Medical University

基　　金：重庆市自然科学基金面上项目(CSTB2022NSCQ-MSX0086,CSTB2024NSCQ-MSX1180);四川省中央引导地方科技发展专项项目(区域创新体系建设-成都-重庆双城经济圈,2023ZYD0283);重庆医科大学药学院2024年“药创之星”本科生科研素养培育计划项目(YCZX20240705)。

摘　　要：目的观察青蒿琥酯(artesunate,AS)对多囊卵巢综合征(polycystic ovary syndrome,PCOS)模型小鼠的治疗作用,并对其作用机制进行初步探讨。方法25只雌性C57BL/6J小鼠按照随机数字表法分为(n=5):对照组(Control组)、模型组(PCOS组)、青蒿琥酯低剂量组(AS15组)、青蒿琥酯高剂量组(AS30组)和二甲双胍组(metformin,Met组)。除Control组外,其余各组小鼠皮下注射脱氢表雄酮(dehydroepiandrosterone,DHEA,60 mg/kg)并给予高脂饲料21 d以构建PCOS模型,AS组腹腔分别注射15和30 mg/kg剂量的青蒿琥酯,Met组灌胃给予200 mg/kg剂量的二甲双胍,每日1次,持续6周。采用ELISA法检测小鼠血清睾酮(testosterone,T)、空腹胰岛素(fasting insulin,FINS)、黄体生成素(luteinizing hormone,LH)、卵泡刺激水平(follicle-stimulating hormone,FSH),并计算LH/FSH比值;采用全自动生化仪检测小鼠空腹血糖(fasting blood glucose,FBG)、甘油三酯(triglyceride,TG)和总胆固醇(total cholesterol,TC)水平,并计算胰岛素抵抗指数(homeostasis model assessment of insulin resistance,HOMA-IR);采用HE染色观察小鼠动情周期,以及卵巢、子宫组织病理形态改变;采用免疫荧光染色观察小鼠卵巢组织中p-eIF2α、ATF4、CHOP蛋白表达情况。体外,使用100μmol/L DHEA诱导人颗粒细胞瘤样细胞系KGN细胞模拟PCOS的高雄激素环境,以5和10μg/mL浓度的AS分别处理24 h后,Western blot检测内质网应激信号通路蛋白表达情况。结果与Control组相比,PCOS组小鼠动情周期紊乱,卵巢出现多囊样改变,血清T、LH/FSH比值显著升高(P<0.05);代谢方面,PCOS小鼠HOMA-IR、TC、TG水平明显升高(P<0.01);PCOS小鼠卵巢颗粒细胞和KGN细胞中p-eIF2α、ATF4、CHOP表达增加(P<0.05)。而与PCOS组相比,AS各剂量组小鼠卵巢和子宫病理形态改变显著改善,血清T、LH/FSH比值也明显降低(P<0.05);代谢方面,AS各剂量组小鼠HOMA-IR、TC、TG水平明显降低(P<0.05);AS各剂量组小鼠卵巢颗粒细胞和KObjective To investigate the therapeutic efficacy of artesunate(AS)on polycystic ovary syndrome(PCOS)in mice and explore the potential mechanism primarily.Methods Twenty-five female C57BL/6J mice were randomly divided into Control group,model group(PCOS group),low-and high-dose AS groups(AS15 and AS30 groups)and metformin group(Met group).In addition to the Control group,the mouse model of PCOS was established by subcutaneous injection of dehydroepiandrosterone(DHEA,60 mg/kg)following by a high-fat diet for 21 d.After modeling,AS of 15 and 30 mg/kg was intraperitoneally injected into the mice of the AS15 and AS30 groups,respectively,and 200 mg/kg Met was given to those of the Met group by gavage,once per day,for 6 weeks.ELISA was used to detect serum testosterone(T),fasting insulin(FINS),luteinizing hormone(LH)and follicle-stimulating hormone(FSH),and the LH/FSH ratio was calculated.The levels of fasting blood glucose(FBG),triglyceride(TG)and total cholesterol(TC)were detected by automatic biochemical analyzer,and the homeostasis model assessment of insulin resistance(HOMA-IR)was calculated.The estrous cycle was observed,and HE staining was performed for pathological changes in the ovary and uterus.Immunofluorescence assay was employed to measure the expression of p-eIF2α,ATF4 and CHOP in the ovarian tissue.After steroidogenic human granulosa-like tumor cell line KGN were exposed to 100μmol/L DHEA to simulate the hyperandrogen environment of PCOS,and then treated with 5 and 10μg/mL AS for 24 h,the protein levels of endoplasmic reticulum stress signaling pathway was detected by Western blotting.Results Compared with the Control group,the PCOS mice had disturbed estrous cycle,polycystic changes in the ovaries,and significantly increased serum T level and LH/FSH ratio(P<0.05),and obviously elevated HOMA-IR,TC and TG levels in terms of metabolism(P<0.01).The expression levels of p-eIF2α,ATF4 and CHOP were notably up-regulated in the ovarian granulosa cells of PCOS mice and KGN cells after DHEA exposure(P<0.05).Ad

关 键 词：青蒿琥酯 多囊卵巢综合征 生殖功能 糖脂代谢 内质网应激 

分 类 号：R282.71[医药卫生—中药学] R285.6[医药卫生—中医学] R711.75