基于AMH/AMHRⅡ信号通路探讨消囊调经汤治疗PCOS-IR模型大鼠的作用机制  

Exploration of Mechanism of Xiaonang Tiaojing Decoction in Treating PCOS-IR Model Rats Based on AMH/AMHRⅡSignaling Pathway

作　　者：周艳艳 袁俊俊[2] 黄旭博 刘沛沛 吴梦瑶 董亚娜 ZHOU Yanyan;YUAN Junjun;HUANG Xubo;LIU Peipei;WU Mengyao;DONG Yana(Henan Province Hospital of Traditional Chinese Medicine/The Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450002,China;Henan University of Chinese Medicine,Zhengzhou 450046,China;Huanghe Science and Technology University,Zhengzhou 450000,China)

机构地区：[1]河南省中医院/河南中医药大学第二附属医院,郑州450002 [2]河南中医药大学,郑州450046 [3]黄河科技学院,郑州450000

出　　处：《中国实验方剂学杂志》2023年第9期137-145,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省科技攻关项目(212102311138)。

摘　　要：目的:基于抗苗勒氏管激素(AMH)/抗苗勒氏管激素Ⅱ型受体(AMHRⅡ)信号通路,探讨消囊调经汤改善多囊卵巢综合征伴胰岛素抵抗(PCOS-IR)模型大鼠的作用机制。方法:48只成年雌性SD大鼠随机分为空白组、模型组、消囊调经汤组(11.4 g·kg^(-1)·d^(-1))、达英-35组(0.21 g·kg^(-1)·d^(-1)),除空白组外各组大鼠通过高脂高糖饮食联合来曲唑构建PCOS-IR模型,给药组灌胃给予相应剂量药物15 d,每给药4 d间隔1 d,空白组及模型组灌胃给予等体积生理盐水。治疗期间每天记录大鼠动情周期,治疗结束后记录体质量、Lee’s指数,采用酶联免疫吸附测定法(ELISA)测定AMH、促黄体生成素(LH)、LH/卵泡刺激素(FSH)、睾酮(T)、雌二醇(E2)水平,血糖仪检测空腹血糖(FPG),放射免疫分析法(RIA)检测空腹胰岛素(FINS)水平,计算胰岛素抵抗指数(HOMA-IR)、胰岛素敏感性指数(QUICKI),全自动生化检测仪测定甘油三酯(TG)、总胆固醇(TC)水平,苏木素-伊红(HE)染色观察卵巢组织形态学变化,免疫组织化学法(IHC)检测卵巢组织内AMHRⅡ、骨形态发生蛋白-15(BMP-15)、Smad5蛋白水平,蛋白免疫印迹法(Western blot)分析AMHRⅡ、BMP-15、Smad5蛋白表达水平。结果:与空白组大鼠比较,模型组大鼠阴道脱落细胞可见大量白细胞,以动情间期为主;体质量、Lee’s指数、糖脂代谢指标(FPG、FINS、HOMA-IR、TG、TC)、AMH及性激素(LH、LH/FSH、T)水平显著上升(P<0.01),QUICKI及E2水平显著下降(P<0.01);卵巢表面囊性突起较多、湿重较大,卵巢组织有较多闭锁状、囊性扩张性卵泡,颗粒细胞层厚度减少,不见卵母细胞;卵巢组织中AMHRⅡ蛋白表达水平显著上升(P<0.01),BMP-15、Smad5蛋白表达水平显著下降(P<0.01)。与模型组大鼠比较,消囊调经汤组大鼠阴道脱落细胞于治疗第5~6天起见角化细胞,逐渐出现稳定的动情周期;体质量、Lee’s指数、糖脂代谢指标(FPG、FINS、HOMA-IR、TG、TC)�Objective:To investigate the mechanism of Xiaonang Tiaojing decoction(XNTJD)in improving polycystic ovary syndrome with insulin resistance(PCOS-IR)model rats based on anti-Müllerian hormone(AMH)/AMH typeⅡreceptor(AMHRⅡ)signaling pathway.Method:Forty-eight adult female SD rats were randomly divided into the blank group,model group,XNTJD group(11.4 g·kg^(-1)·d^(-1))and Diane-35 group(0.21 g·kg^(-1)·d^(-1)),PCOS-IR model was established by high-fat and high-sugar diet combined with letrozole in rats of all groups except the blank group,rats in the administration groups were given the corresponding dose of drugs by gavage for 15 days with an interval of 1 d every 4 d,normal saline of the same volume was given to the blank group and the model group.Estrous cycle was recorded daily during treatment.At the end of treatment,body weight and Lee's index were recorded,AMH,luteinizing hormone(LH),LH/follicle stimulating hormone(FSH),testosterone(T)and estradiol(E2)levels were measured by enzyme-linked immunosorbent assay(ELISA),fasting plasma glucose(FPG)was measured by glucometer,fasting insulin(FINS)level was measured by radioimmunoassay(RIA),and the insulin resistance index(HOMA-IR)and insulin sensitivity index(QUICKI)were calculated,triglyceride(TG)and total cholesterol(TC)levels were measured by automatic biochemical analyzer,hematoxylin-eosin(HE)staining was used to observe the morphological changes of the ovary,the levels of AMHRⅡ,bone morphogenetic protein-15(BMP-15)and Smad5 in ovarian tissues were detected by immunohistochemistry(IHC),Western blot was used to analyze the protein expression levels of AMHRⅡ,BMP-15 and Smad5.Result:Compared with the blank group,a large number of leukocytes were observed in the vaginal exfoliated cells of rats in the model group,mainly in diestrus,the levels of body weight,Lee's index,glucose-lipid metabolism indexes(FPG,FINS,HOMA-IR,TG,TC),AMH and sex hormones(LH,LH/FSH,T)were significantly increased(P<0.01),and QUICKI and E2 levels were significantly decreased(P<0.01),th

关 键 词：多囊卵巢综合征(PCOS) 胰岛素抵抗(IR) 消囊调经汤 抗苗勒氏管激素(AMH)/抗苗勒氏管激素Ⅱ型受体(AMHRⅡ)信号通路 糖脂代谢 骨形态发生蛋白-15(BMP-15) SMAD5 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学] R96R588