Wu Yuanyuan, Long Yunxia, Su Guangheng, Fan Xiangping, He Guozhen, Luo Zhijuan, Luo Songping
First School of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangdong Province, Guangzhou City, 510405, China.
Department of Gynecology, Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, Guangxi Province, Nanning City, 530011, China.
Comb Chem High Throughput Screen. 2025;28(4):724-736. doi: 10.2174/0113862073291139240506114446.
Premature Ovarian Insufficiency (POI) is a disease suffered by women under the age of 40 when ovarian function has declined, seriously affecting both the physical and mental health of women. Guiluoshi Anzang decoction (GLSAZD) has been used for a long time and has a unique therapeutic effect on improving ovarian function. This study aims to investigate the mechanism of GLSAZD in treating POI through network pharmacology, molecular docking, and experimental verification.
In this study, the active ingredients of Guiluoshi Anzang Decoction and the targets of POI were obtained from TCMSP, BATMANN-TCM, Uniprot, GeneCards, and other databases, and network pharmacology analysis was performed. Molecular docking was conducted to validate the affinity of the main active ingredient of GLSAZD to key POI targets. A POI SD rat model was established, and HE staining, ELISA, Real-time PCR, and Western blot experiments were performed to verify the predicted core targets and the therapeutic effects.
10 core targets and the top 5 ingredients were screened out. Molecular docking showed core targets AKT1, CASP3, TNF, TP53, and IL6 had stable binding with the core 5 ingredients quercetin, kaempferol, beta-sitosterol, luteolin, and Stigmasterol. GO and KEGG enrichment analysis demonstrated the mechanism involved in the positive regulation of gene expression, PI3K-AKT signaling pathway, and apoptosis signaling pathways. Animal experiments indicated GLSAZD could up-regulate the protein expression of p-PI3K and p-AKT1 and the mRNA expression of STAT3 and VEGF, down-regulate TP53 and Cleaved Caspase-3 protein expression in rat`s ovarian tissues and serum TNF-α and IL-6 protein levels, activate PI3K-AKT signaling pathway and inhibit the apoptosis signaling pathway.
GLSAZD treats POI through multi-component, multi-target, and multi-pathway approaches. This study provided evidence for its clinical application in treating POI and shed light on the study of traditional medicine of the Guangxi Zhuang Autonomous Region in China.
卵巢早衰（POI）是40岁以下女性卵巢功能减退所患疾病，严重影响女性身心健康。归蒌氏安脏汤（GLSAZD）长期应用，对改善卵巢功能有独特疗效。本研究旨在通过网络药理学、分子对接和实验验证探讨GLSAZD治疗POI的机制。
本研究从中药系统药理学数据库与分析平台（TCMSP）、中药分子机制生物信息学数据库（BATMANN-TCM）、通用蛋白质数据库（Uniprot）、基因卡片数据库（GeneCards）等数据库获取归蒌氏安脏汤的活性成分及POI的靶点，并进行网络药理学分析。进行分子对接以验证GLSAZD主要活性成分与POI关键靶点的亲和力。建立POI SD大鼠模型，进行苏木精-伊红（HE）染色、酶联免疫吸附测定（ELISA）、实时荧光定量聚合酶链反应（Real-time PCR）和蛋白质免疫印迹法（Western blot）实验，以验证预测的核心靶点及治疗效果。
筛选出10个核心靶点和前5种成分。分子对接显示核心靶点蛋白激酶B1（AKT1）、半胱天冬酶3（CASP3）、肿瘤坏死因子（TNF）、肿瘤蛋白53（TP53）和白细胞介素6（IL6）与核心5种成分槲皮素、山柰酚、β-谷甾醇、木犀草素和豆甾醇有稳定结合。基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析表明其机制涉及基因表达的正调控、磷脂酰肌醇-3激酶-蛋白激酶B（PI3K-AKT）信号通路和凋亡信号通路。动物实验表明GLSAZD可上调大鼠卵巢组织中磷酸化磷脂酰肌醇-3激酶（p-PI3K）和磷酸化蛋白激酶B1（p-AKT1）的蛋白表达以及信号转导和转录激活因子3（STAT3）和血管内皮生长因子（VEGF）的mRNA表达，下调TP53和裂解的半胱天冬酶3（Cleaved Caspase-3）蛋白表达以及血清肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）蛋白水平，激活PI3K-AKT信号通路并抑制凋亡信号通路。
GLSAZD通过多成分、多靶点、多途径治疗POI。本研究为其治疗POI的临床应用提供了依据，并为中国广西壮族自治区传统医学的研究提供了思路。
