基于生物信息学技术探讨左归丸治疗多囊卵巢综合征的机制  

Bioinformatics Reveals Mechanism of Zuoguiwan in Treating Polycystic Ovary Syndrome

作　　者：张金融 李浩天 黄鸿铭 邓阿黎 赵敏 ZHANG Jinrong;LI Haotian;HUANG Hongming;DENG Ali;ZHAO Min(School of Basic Medical Sciences,Hubei University of Chinese Medicine,Wuhan 430065,China;Affiliated Hospital of Hubei University of Chinese Medicine,Wuhan 430000,China)

机构地区：[1]湖北中医药大学基础医学院,武汉430065 [2]湖北中医药大学附属医院,武汉430000

出　　处：《中国实验方剂学杂志》2024年第17期77-86,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：湖北省自然科学基金计划项目(2023AFD114)。

摘　　要：目的:该研究旨在利用现代网络药理学技术与液相色谱-质谱联用仪(LC-MS)代谢组学技术探讨左归丸改善多囊卵巢综合征(PCOS)的潜在机制。方法:该研究基于左归丸的活性成分及其治疗多囊卵巢综合征的潜在靶点进行生物信息学方法预测,将SD大鼠分为空白组(Con),PCOS模型制备组,采用来曲唑(1 mg·kg^(-1))灌胃+高脂饮食(HFD),建立PCOS大鼠模型并鉴定。造模结束后将PCOS模型制备组分为模型组(Mod)灌胃等量生理盐水、二甲双胍组(Met)灌胃300 mg·kg^(-1)二甲双胍,左归丸组(Zgw)灌胃左归丸浓缩液(1.62 g·kg^(-1)),记录体质量、发情周期,苏木素-伊红染色观察卵巢形态,酶联免疫吸附测定法(ELISA)测定血清促卵泡激素(FSH)、黄体生成素(LH)、抗苗勒管激素(AMH)、睾酮(T)、雌二醇(E_(2))含量,计算LH/FSH比值,通过正交偏最小二乘法判别分析(OPLS-DA)对大鼠血清代谢组学进行分析,筛选代谢物的富集通路,联合网络药理学,进行关联分析,构建化合物-反应-酶-基因网络。结果:数据库筛选出左归丸的503种潜在靶蛋白,以及5843个多囊卵巢综合征的疾病基因,交集靶点共271个。基因本体(GO)富集分析涉及对脂多糖的反应等,京都基因和基因组百科全书(KEGG)富集得到119条通路。动物实验发现与Con组比较,Mod组体质量、LH、AMH水平显著升高(P<0.01),FSH水平差异无统计学意义,但LH/FSH比Con组显著升高(P<0.01),雌激素水平显著下降(P<0.01),囊性卵泡增多,与Mod组比较,Zgw组与Met组体质量显著下降(P<0.01),Zgw组大鼠闭锁卵泡和囊性卵泡减少,成熟卵泡和黄体增多,颗粒层变厚。Zgw组大鼠血清T、FSH、LH、AMH、LH/FSH水平显著下降(P<0.01),经左归丸治疗后,E_(2)水平显著升高(P<0.01)。代谢组学PCA与OPLSDA结果显示Zgw组与Mod组之间差异有统计学意义,代谢差异物中,Zgw组中有26个代谢物上调,网络药理学KEGG富集通路与代谢物富集通路共有8条�Objective:To explore the potential mechanism of Zuoguiwan in ameliorating polycystic ovary syndrome(PCOS)by network pharmacology and liquid chromatography-mass spectrometry(LCMS)-based metabolomics.Method:The active ingredients and potential targets of Zuoguiwan for treating PCOS were predicted by bioinformatics.SD rats were assigned into a control group and a modeling group.The rat model of PCOS was established by gavage with letrozole(1 mg·kg^(-1))combined with feeding with a high-fat diet.At the end of modeling,the modeled rats were assigned into model(normal saline),metformin(300 mg·kg^(-1)),and Zuoguiwan(concentrate 1.62 g·kg^(-1))groups.The body weight and oestrous cycle of each rat were recorded,and the ovary was stained with hematoxylin and eosin for observation of ovarian morphology.Enzyme-linked immunosorbent assay was employed to determine the serum levels of follicle-stimulating hormone(FSH),luteinizing hormone(LH),anti-mullerian hormone(AMH),testosterone(T),and estradiol(E_(2)),and the LH/FSH ratio was calculated.Serum metabolomics of rats was conducted by orthogonal partial least squares-discriminant analysis(OPLS-DA)to screen the metabolite-enriched pathways.Furthermore,network pharmacology and association analysis were employed construct the compound-response-enzyme-gene network.Result:A total of 503 potential targets of Zuoguiwan and 5843 targets of PCOS were screened out,with 271 common targets.The Gene Ontology enrichment analysis revealed that the common targets were involved in the response to lipopolysaccharide,etc.,and the Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment yielded 119 pathways.Animal experiments showed that compared with the control group,the model group presented increased body weight(P<0.01),elevated LH and AMH levels(P<0.01),increased LH/FSH ratio(P<0.01),lowered E_(2) level(P<0.01),and increased cystic follicles.Compared with the model group,Zuoguiwan and metformin decreased the body weight(P<0.01),reduced atretic follicles and cystic follicles,increased mature

关 键 词：左归丸 多囊卵巢综合征 网络药理学 代谢组学 类固醇激素生物合成途径 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33