补肾调肝方对早发性卵巢功能不全合并失眠大鼠卵巢功能及睡眠的影响  

The impact of Bushen Tiaogan formula on ovarian function and sleep in rats with premature ovarian insufficiency combined with insomnia

作　　者：郑涵予 刘雁峰[1] 张泽灵 金凡惠 周雨玫 徐子涵 ZHENG Hanyu;LIU Yanfeng;ZHANG Zeling;JIN Fanhui;ZHOU Yumei;XU Zihan(Dongzhimen Hospital of Beijing University of Chinese Medicine,Beijing 100700,China;不详)

机构地区：[1]北京中医药大学东直门医院妇科,100700 [2]北京中医药大学第三附属医院妇科

出　　处：《环球中医药》2023年第12期2444-2450,共7页Global Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82074480)。

摘　　要：目的观察补肾调肝方对雷公藤多苷联合对氯苯丙氨酸(4-chloro-DL-phenylalanine,PCPA)诱导的早发性卵巢功能不全(premature ovarian insufficiency,POI)合并失眠大鼠模型的影响,探讨其作用机制。方法选取具有规律动情周期的雌性SD大鼠,随机分为模型组(40只)、经典模型组(8只)和空白组(8只),经典模型组通过雷公藤多苷片造模,模型组通过雷公藤多苷片联合PCPA构建POI合并失眠大鼠模型,造模成功后将模型大鼠随机分为模型组、西药组、褪黑素组、中药高剂量组、中药低剂量组,每组8只,分别予药物干预20天。采用阴道涂片观察动情周期;苏木精—伊红(hematoxylin-eosin,HE)染色观察卵巢组织;戊巴比妥钠睡眠实验观察睡眠变化;酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测血清促卵泡激素(follicle-stimulating hormone,FSH)、雌激素(luteinizing hormone,E2)、5-羟色胺(5-hydroxyryptamine,5-HT);蛋白质印迹法检测卵巢细胞色素P450芳香化酶(cytochrome P450arom,Cyp19a1)蛋白表达量;荧光定量逆转录聚合酶链式反应法检测卵巢雌激素受体β(estrogen receptorβ,ERβ)、卵泡刺激素受体(FSH receptor,FSHR)、Cyp19a1的mRNA含量。结果与模型组比较,各治疗组大鼠动情周期明显改善,发育中卵泡增多,睡眠潜伏期均显著缩短(P<0.05),睡眠时间不同程度延长,FSH降低,E2、5-HT升高(P<0.05),卵巢Cyp19a1蛋白表达及相应mRNA含量升高(P<0.05),卵巢ERβmRNA、FSHR mRNA含量升高(P<0.05);与西药组相比,中药高剂量组睡眠时间延长(P<0.05),5-HT升高(P<0.05),Cyp19a1 mRNA、FSHR mRNA升高(P<0.05);与褪黑素组相比,中药高剂量组睡眠潜伏期缩短(P<0.05),E2水平升高(P<0.05),FSH降低(P<0.05),Cyp19a1及mRNA及ERβmRNA、FSHR mRNA均升高(P<0.05)。结论补肾调肝方有改善POI合并失眠大鼠卵巢功能和睡眠的作用,可调节性激素和神经递质水平,其作用机制可能与增加卵巢组织ERβ、FSHR、Cyp19a1表达,促进Objective To observe the effect of Bushen Tiaogan formula on the combined model of premature ovarian insufficiency(POI)and insomnia induced by Tripterygium Glycosides combined with 4-chloro-DL-phenylalanine(PCPA)in rats,and to explore its underlying mechanism.Methods Female SD rats with regular estrous cycles were randomly divided into model group(40 rats),classic model group(8 rats),and blank group(8 rats).The classic model group was modeled using Tripterygium Glycosides,while the model group was built by combining Tripterygium Glycosides with PCPA to induce the combined model.After modeling was successful,the model rats were randomly divided into the model group,western medicine group,melatonin group,high-dose Chinese medicine group,and low-dose Chinese medicine group,with 8 rats in each group.Drug intervention was given for 20 days.Vaginal smears were used to observe the estrous cycle.Hematoxylin-eosin(HE)staining was used to observe ovarian tissue.Pentobarbital sodium sleep experiment was used to observe sleep changes.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum follicle-stimulating hormone(FSH),estrogen(E2),and 5-hydroxyryptamine(5-HT).Protein immunoblotting was used to detect the protein expression of cytochrome P450arom(Cyp19a1)in ovarian cells.Fluorescence quantitative reverse transcription polymerase chain reaction(RT-PCR)was used to detect the mRNA content of estrogen receptor beta(ERβ),follicle-stimulating hormone receptor(FSHR),and Cyp19a1 in the ovary.Results Compared with the model group,the estrous cycle in each treatment group of rats were improved significantly,with an increase in developing follicles.The latent period of sleep was significantly decreased(P<0.05),and sleep time was prolonged to varying degrees.FSH decreased,while E2 and 5-HT increased(P<0.05).The protein expression and corresponding mRNA content of Cyp19a1 in the ovary were increased(P<0.05),and the mRNA content of ovarian ERβand FSHR increased(P<0.05).Compared with the western medicine group,the high-dose C

关 键 词：早发性卵巢功能不全 补肾调肝方 卵巢功能 睡眠 性激素 

分 类 号：R285.5[医药卫生—中药学]