补肾调肝方对卵巢储备功能下降合并慢性心理应激大鼠卵巢功能的影响及机制研究  

Mechanism and effects of Bushen Tiaogan Formula in diminished ovarian reserve rats with chronic psychological stress

作　　者：刘柳青 刘雁峰[2] 王悦竹[3] 郑涵予 金凡惠 LIU Liu-qing;LIU Yan-feng;WANG Yue-zhu;ZHENG Han-yu;JIN Fan-hui(Traditional Chinese Medicine College of Anhui University of Chinese Medicine,Hefei 230012,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China;Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]安徽中医药大学中医学院,合肥230012 [2]北京中医药大学东直门医院,北京100700 [3]北京中医药大学,北京100029

出　　处：《中华中医药杂志》2022年第8期4459-4465,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：北京中医药大学东直门医院横向课题(No.HX-DZM-2019010);中央高校基本科研业务费专项资金(No.2019-JYB-XS-148)。

摘　　要：目的:探讨补肾调肝方改善卵巢功能的可能机制。方法:将80只动情周期正常的雌性SD大鼠随机分为对照组(20只)和造模组(60只),造模组运用雷公藤多苷片联合慢性不可预知温和刺激法建立卵巢储备功能下降(DOR)疾病模型。造模后随机分为模型组,中药高、中、低剂量组和西药组。中药高、中、低剂量组分别予高、中、低剂量的补肾调肝方干预,西药组予雌孕激素序贯法干预,均干预20 d。观察动情周期,ELISA法检测血清抗苗勒管激素(AMH)水平,HE染色观察卵巢组织形态学表现,Western Blot法检测卵巢PI3K、AKT、mTOR、p70S6K、CyclinD2、p27^(Kip1)蛋白表达量,qRT-PCR法检测上述蛋白相应mRNA含量。结果:与对照组比较,模型组大鼠动情周期紊乱,血清AMH水平显著下降(P<0.05),闭锁卵泡增加,发育中卵泡减少;卵巢PI3K、AKT、mTOR、p70S6K、CyclinD2蛋白及相应mRNA表达显著下降(P<0.05),p27^(Kip1)蛋白及mRNA表达显著升高(P<0.05)。与模型组比较,各治疗组卵巢组织形态学表现有不同程度恢复,西药组动情周期显著改善(P<0.05),各药物干预组AMH水平均显著升高(P<0.05);各药物干预组PI3K、AKT、mTOR、p70S6K、CyclinD2蛋白表达显著升高(P<0.05),中药高剂量组及西药组p27^(Kip1)蛋白及mRNA表达显著下降(P<0.05),中药高剂量组PI3K、p70S6K、CyclinD2,中药中剂量组mTOR、CyclinD2,中药低剂量组p70S6K,西药组AKT、CyclinD2 mRNA表达均显著升高(P<0.05)。结论:补肾调肝方可有效改善DOR大鼠卵巢储备,其作用机制可能与调节PI3K/AKT信号通路及其下游因子mTOR、p70S6K、CyclinD2、p27^(Kip1)蛋白表达,从而改善细胞生长有关。Objective:To investigate the possible mechanism of Bushen Tiaogan Formula (BTF) in improving ovarian function.Methods:A total of 80 female Sprague-Dawley (SD) rats with regular estrous cycles were selected and randomly divided into control group (20 rats) and model group (60 rats).The model of diminished ovarian reserve (DOR) was established by gavage of tripterygium glycosides combined with chronic unpredictable mild stress.After modeling,model rats were divided into model group,high dose Chinese medicine group,medium dose Chinese medicine group,low dose Chinese medicine group and western medicine group at random.The Chinese medicine groups were treated with high,medium and low concentration of BTF respectively,and the western medicine group was treated with estrogen progesterone sequence.The treatment lasted20 days.The observation indexes included estrous cycles,serum AMH levels detected by ELISA,ovarian histomorphology appearance stained by hematoxylin eosin (HE),expressions and contents of PI3K,AKT,mTOR,p70S6K,CyclinD2,p27^(Kip1)and their corresponding mRNAs in ovary detected by Western Blot and qRT-PCR.Results:Compared with control group,in model group,the number of rats with abnormal estrous cycle increased and serum AMH level decreased (P<0.05),atretic follicles increased and developing follicles decreased,and the expressions of PI3K,AKT,mTOR,p70S6K,CyclinD2 and their mRNAs in ovary decreased,while the expressions of p27^(Kip1) and p27^(Kip1) mRNA increased (P<0.05).Compared with model group,ovarian histomorphology appearance in treatment groups recovered to different degrees,and estrous cycle in west medicine group and AMH levels in each treatment group were improved (P<0.05).The protein expressions of PI3K,AKT,mTOR,p70S6K,CyclinD2 in each treatment group increased significantly (P<0.05).The protein expression and mRNA content of p27^(Kip1) in high dose Chinese medicine group and western medicine group decreased significantly (P<0.05).The mRNA contents increased significantly,including PI3K,p70S6K and Cyc

关 键 词：卵巢储备功能下降 补肾调肝方 PI3K/AKT信号通路 蛋白合成 细胞周期 细胞生长 

分 类 号：R285.5[医药卫生—中药学]