补肾调肝方对卵巢储备功能下降大鼠卵巢/海马功能及PI3K/AKT/mTOR信号通路的影响  

Effects of Bushen Tiaogan formula on ovarian and hippocampal function and PI3K/AKT/mTOR signal pathway in rats with diminished ovarian reserve

作　　者：刘柳青 刘雁峰[1] 潘雪 许梦白 吴豆豆 杨绿 LIU Liuqing;LIU Yanfeng;PAN Xue;XU Mengbai;WU Doudou;YANG Lyu(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学东直门医院妇科,100700

出　　处：《环球中医药》2021年第5期822-829,共8页Global Traditional Chinese Medicine

基　　金：中央高校基本科研业务费专项资金资助(2019-JYB-XS-148)。

摘　　要：目的观察补肾调肝方对卵巢储备功能下降大鼠卵巢/海马功能的调节作用,探讨其对PI3K/AKT/mTOR信号通路的调控作用。方法选取具有规律动情周期的雌性SD大鼠,利用随机数字表随机分为模型组和对照组。模型组采取雷公藤多苷片(75 mg/kg/d)联合慢性不可预知温和刺激法造模。造模成功后随机分为模型组、中药高剂量组、中药中剂量组、中药低剂量组、西药组。中药组予不同浓度补肾调肝方(5.46 g/kg/d,2.73 g/kg/d,1.365 g/kg/d)治疗,西药组予戊酸雌二醇片(0.105 mg/kg/d)、地屈孕酮片(2.1 mg/kg/d)序贯治疗,共计20天。观察指标包括旷场试验、卵巢组织苏木精—伊红染色、酶联免疫吸附法检测海马神经递质5-羟色胺及多巴胺、蛋白质印迹法检测卵巢及海马组织PI3K/AKT/mTOR通路蛋白的表达及活化情况、荧光定量逆转录聚合酶链式反应法检测相应mRNA含量表达。结果与对照组相比,模型组大鼠活动次数减少(P<0.05),海马神经递质含量下降(P<0.01),卵巢闭锁卵泡增加,黄体和血管减少,卵巢、海马组织通路蛋白表达、活化及相应mRNA含量下降(均P<0.01)。与模型组相比,各治疗组大鼠活动次数有不同程度的增加,神经递质含量明显升高(P<0.01),卵巢中黄体、血管丰富,有各级正常卵泡;卵巢和海马组织通路蛋白的表达、活化及相应mRNA的表达有不同程度改善,中药高剂量组改善最明显(均P<0.01)。结论补肾调肝方能显著改善卵巢储备功能下降合并慢性心理应激大鼠的卵巢储备和海马功能,其作用机制可能与上调PI3K/AKT/mTOR信号通路有关。Objective To observe the regulation effect of Bushen Tiaogan formula on ovarian and hippocampal function in rats with diminished ovarian reserve,and to explore the regulation effect of Bushen Tiaogan formula on PI3K/AKT/mTOR signal pathway.Methods Female Sprague-Dawley(SD)rats with regular estrous cycles were selected and randomly divided into model group and control group using a random number table.The rat model was established by giving a gavage of tripterygium glycosides(75 mg/kg/d)combined with chronic unpredictable mild stress.After successful modeling,they were randomly divided into model group,high,medium and low dose of Chinese medicine group,and western medicine group.Chinese medicine groups were treated with different concentrations of Bushen Tiaogan formula(5.46 g/kg/d,2.73 g/kg/d,1.365 g/kg/d).Western medicine group was given sequential treatment with estradiol valerate tablets(0.105 mg/kg/d)and didrogesterone tablets(2.1 mg/kg/d),and the treatment lasted 20 days.The observation indexes included open field test,HE staining of ovarian tissue,5-hydroxytryptamine(5-HT)and dopamine(DA)in hippocampal neurotransmitters detected by ELISA,protein expression and activation of PI3K/Akt/mTOR pathway in ovarian and hippocampal tissues detected by Western Blot,and the corresponding mRNA contents detected by qRT-PCR.Results Compared with the control group,the rats in the model group showed less activity times(P<0.05),lower contents of neurotransmitter in hippocampal(P<0.01),more ovarian atresia follicles,and less corpus luteum and blood vessels,less proteins expression,activation and corresponding mRNA contents on PI3K/AKT/mTOR signal pathway in ovary and hippocampus(P<0.01).Compared with model group,the activity times in the treatment group rats increased to different degrees,and the content of neurotransmitters increased significantly(P<0.01),corpus luteum and blood vessels are rich,there are all levels of normal follicles.The expression and activation of pathway proteins in ovary and hippocampus and the corresp

关 键 词：补肾调肝方 卵巢储备功能下降 慢性心理应激 PI3K/AKT/mTOR信号通路 海马功能 卵巢功能 

分 类 号：R285.5[医药卫生—中药学]