摘要
[目的]运用4-去氧乙烯基环已烯(VCD)建立早发性卵巢功能不全(POI)小鼠模型,探究乌鸡白凤片对VCD致POI小鼠的治疗作用。[方法]1)建立POI小鼠模型:8周龄雌性昆明小鼠60只,随机分为正常(CON)组30只,模型(MOD)组30只。MOD组腹腔注射160 mg/(kg·d)VCD复制POI模型,CON组注射同体积芝麻油。造模期间每日阴道涂片观察动情周期;造模第15~65天,每5 d进行眼眶取血,酶联免疫吸附(ELISA)法检测血清促卵泡生长素(FSH)、促黄体生成素(LH)水平;造模第45~65天,每5 d取小鼠子宫卵巢,称质量后计算脏器指数,苏木精-伊红(HE)染色观察卵巢形态学改变。2)乌鸡白凤片对POI小鼠的治疗作用:按照上述造模方法再选取8周龄雌性昆明小鼠60只,造模成功后随机分为MOD组、戊酸雌二醇(E2)组[0.3 mg/(kg·d)]、乌鸡白凤片低、中、高剂量组[410、820、1640 mg/(kg·d)],每组10只。CON组和MOD组予以等体积羧甲基纤维素钠(CMC-Na)灌胃。每日1次,连续给药45 d,给药期间观察小鼠动情周期。于给药第46天处死小鼠,ELISA法检测血清FSH、LH水平,取小鼠子宫卵巢,称质量后计算脏器指数,HE染色观察卵巢形态学改变。[结果]1)小鼠POI模型的建立:与CON组相比,MOD组小鼠第55天动情周期紊乱持续4周,VCD造模第50、55天MOD组小鼠血清FSH、LH显著升高2次,子宫、卵巢湿质量及脏器指数显著下降,生长卵泡数量显著减少。2)乌鸡白凤片对POI小鼠的治疗作用:灌胃给药45 d后,与CON组相比,MOD组小鼠动情周期紊乱,血清FSH、LH显著升高,子宫、卵巢湿质量及脏器指数显著下降,生长卵泡数量显著减少。与MOD组相比,乌鸡白凤片中、高剂量组小鼠动情周期全部恢复正常,且乌鸡白凤片各剂量组小鼠血清FSH、LH下降,子宫、卵巢湿质量及脏器指数显著升高,卵巢内原始卵泡数量明显增多。[结论]160 mg/(kg·d)VCD腹腔注射15 d后,观察40 d,总计造模55 d后可制备出POI小鼠模型;
[Objective]Establish the mouse model of primary ovarian insufficiency(POI)with 4-deoxyvinylcyclohexene(VCD)and explore the therapeutic effect of Wuji Baifeng Tablets on VCD-induced POI mice.[Methods]1)The POI mouse model was established:sixty 8 week old Kunming female mice were randomly divided into 30 control(CON)mice and 30 model(MOD)mice.In MOD group,the POI model was reproduced by intraperitoneal injection of 160 mg/(kg·d)VCD,and CON was injected with sesame oil of the same volume.During the modeling period,the estrus cycle was observed by vaginal smear every day;from day 15 to day 65 of modeling,blood was taken from orbit every 5 days,and serum follicle stimulating hormone(FSH)and luteinizing hormone(LH)levels were detected by ELISA.At 45 to 65 days after modeling,the uterus and ovary of mice were taken every 5 days,and the organ index was calculated after weighing,and morphological changes in the ovaries were observed by hematoxylin-eosin(HE)staining.2)Therapeutic effect of Wuji Baifeng Tablets on POI mice:Another sixty 8 week old female Kuming mice were selected according to the above modelling method.After successful modeling,they were randomly divided into MOD group,estradiol valerate(E2)group[0.3 mg/(kg·d)],and low,medium and high dose Wuji Baifeng Tablets groups[410,820,1640 mg/(kg·d)],with 10 mice in each group.Once daily for 45 consecutive days,the estrus cycle of mice was observed during administration.The mice were killed on day 46 of administration,and serum FSH and LH levels were detected by ELISA.The uterus and ovaries of mice were taken,and the viscera index was calculated after weighing.The morphological changes in ovaries were observed by HE staining.[Results]1)Establishment of mouse POI model:compared to the CON group,estrous cycle disorder in the MOD group lasted for 4 weeks on day 55.At day 50 and day 55 of VCD modeling,serum FSH and LH in MOD mice increased significantly twice.The wet weight and organ index of uterus and ovary decreased significantly,and the number of growing follicles
作者
陆兴玥
王娜
高盼微
袁庆
苗琳
张晗
王怡
柴丽娟
LU Xingyue;WANG Na;GAO Panwei;YUAN Qing;MIAO Lin;ZHANG Han;WANG Yi;CHAI Lijuan(Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae,Ministry of Education,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)
出处
《天津中医药》
CAS
2023年第6期764-774,共11页
Tianjin Journal of Traditional Chinese Medicine
基金
国家自然科学基金项目(81102860)
国家中医药管理局中医临床疗效提升项目
天津中新药业集团股份有限公司开发项目(HX2021-47)。