补肾生精调和气血法调控Bcl-2相关线粒体凋亡信号通路改善大鼠卵巢储备功能的研究  

Study on the Bushen Shengjing Tiaohe Qixue Therapy Regulating Bcl-2-related Mitochondrial Apoptotic Signaling Pathway for Improving Ovarian Reserve Function in Rats

作　　者：黄佳梅 蔡紫璨 张花[1,2] 黎玲 叶朝阳 金志春[1,2] HUANG Jia-Mei;CAI Zi-Can;ZHANG Hua;LI Ling;YE Chao-Yang;JIN Zhi-Chun(Hubei University of Chinese Medicine,Wuhan 430065 Hubei,China;Maternal and Child Healthcare Hospital of Hubei Province,Tongji Medical School,Hubei University of Chinese Medicine,Wuhan 430070 Hubei,China)

机构地区：[1]湖北中医药大学,湖北武汉430065 [2]华中科技大学同济医学院附属湖北妇幼保健院,湖北武汉430070

出　　处：《广州中医药大学学报》2023年第1期155-162,共8页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：湖北省卫健委中医药科研项目(重点项目)(编号:ZY2021Z013)。

摘　　要：【目的】探讨补肾生精调和气血法对卵巢储备功能减退(DOR)模型大鼠的干预机制。【方法】将60只性成熟雌性SD大鼠随机分为正常组,模型组,西药组,中药低、中、高剂量组,每组10只。除正常组,其余各组大鼠给予单次腹腔注射环磷酰胺构建DOR模型。于造模次日开始,西药组给予戊酸雌二醇0.105 mg·kg^(-1)·d^(-1)灌胃,中药低、中、高剂量组分别对应给予补肾生精调和气血法中药复方8.075、16.15、32.3 g·kg^(-1)·d^(-1)灌胃,正常组、模型组给予等体积去离子水灌胃,每日1次,连续21 d。给药结束后,采用酶联免疫吸附分析(ELISA)检测血清卵泡刺激素(FSH)、黄体生成素(LH)水平,采用电化学发光法检测血清雌二醇(E2)、抗苗勒管激素(AMH)水平,采用Western Blot法检测卵巢组织血红素氧合酶1(HO-1)、醌氧化还原酶1(NQO-1)、B淋巴细胞瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸蛋白酶3(Caspase-3)和半胱氨酸蛋白酶9(Caspase-9)的蛋白表达水平,采用实时荧光定量聚合酶链反应(PCR)法检测卵巢组织HO-1、NQO-1、Bcl-2、Bax、Caspase-3、Caspase-9的mRNA表达水平。【结果】与正常组比较,模型组血清FSH、LH水平升高,E2、AMH水平降低,卵巢组织HO-1、NQO-1、Bcl-2蛋白表达水平下调,Bax、cleaved Caspase-3、Caspase-9、cleaved Caspase-9蛋白表达水平上调,Caspase-3、Caspase-9 mRNA表达水平下调(均P<0.05);与模型组比较,西药组和中药低、中、高剂量组上述各指标得到一定程度的改善,且中药各剂量组各指标的改善效果与西药组相当。【结论】补肾生精调和气血法通过调控Bcl-2相关线粒体凋亡信号通路,抑制卵巢颗粒细胞凋亡,改善大鼠卵巢储备功能。Objective To investigate the intervention mechanism of Bushen Shengjing Tiaohe Qixue Therapy(a herbal compound prescription mainly with the actions of tonifying the kidney to produce essence qi,and harmonizing qi and blood)on the rats model with diminished ovarian reserve(DOR).Methods Sixty sexually mature female SD rats were randomly divided into normal group,model group,western medicine group,and Chinese medicine low-,medium-and high-dose groups,with 10 rats in each group.Except for the normal group,the DOR model was prepared by single intraperitoneal injection of cyclophosphamide into all the groups.From the next day after modeling,the western medicine group was given Estradiol Valerate at 0.105 mg·kg^(-1)·d^(-1)by gavage,while the Chinese medicine low-,medium-and high-dose groups were given Chinese herbal compound with Bushen Shengjing Tiaohe Qixue Therapy at 8.075,16.15 and 32.3 g·kg^(-1)·d^(-1)by gavage,respectively.The normal group and the model group were given equal volume of deionized water by gavage.All the treatments performed once a day and lasted for consecutive 21 days.After administration,serum follicle stimulating hormone(FSH)and luteinizing hormone(LH)levels were detected by enzyme-linked immunosorbent assay(ELISA),serum estradiol(E2)and anti-mullerian hormone(AMH)levels were detected by electrochemiluminescence assay,the protein relative expressions of heme oxygenase 1(HO-1),quinone oxidoreductase 1(NQO-1),B lymphocytoma 2(Bcl-2),Bcl-2-associated X protein(Bax),cysteine proteinase3(Caspase-3)and cysteine proteinase 9(Caspase-9)in ovarian tissue were detected by Western Blot,the mRNA relative expressions of HO-1,NQO-1,Bcl-2,Bax,Caspase-3,Caspase-9 in ovarian tissue were detected by real time quantitative polymerase chain reaction(PCR).Results Compared with the normal group,the serum FSH and LH levels were increased,E2and AMH levels were decreased,the protein expressions of HO-1,NQO-1and Bcl-2 in ovarian tissue were down-regulated,the protein expressions of Bax,cleaved Caspase-3,Caspase-9 and

关 键 词：补肾生精调和气血法 卵巢储备功能减退 线粒体信号通路 颗粒细胞凋亡 大鼠 

分 类 号：R285.5[医药卫生—中药学]