Aim: To verify the effects of modified Gengnianchun formula (MGNC), a traditional Chinese medicine, on a stressed diminished ovarian reserve (DOR) animal model and predict the underlying mechanisms through network pharmacology strategies.
Methods: Sexually mature female C57BL/6 mice were allocated to five groups, abbreviated as the control (C) group, stress manipulated model (M) group, stress with normal saline gavage (N) group, stress with low-dose MGNC gavage (L) group, and stress with high-dose MGNC gavage (H) group. Body weight and the estrous cycle were monitored during the stress and gavage process. Serum stress hormones and reproductive hormones were evaluated by ELISA. Ovarian follicle counts were calculated, and ovarian follicle-stimulating hormone receptor (FSHR) and anti-Müllerian hormone (AMH) expression were assessed by Western blotting and immunohistochemistry. Network pharmacology strategies included active compound screening, drug and disease target analysis, gene ontology analysis, pathway analysis, and visualization of results.
Results: MGNC treatment significantly decreased serum corticosterone (CORT) and follicle-stimulating hormone (FSH) levels and increased testosterone (T) levels in the H group compared with the M and N groups. Primordial and preantral follicle counts and ovarian AMH and FSHR expression were significantly increased in the H group compared to those in the M and N groups. Through pharmacokinetic screening, we found 244 active compounds in MGNC. A total of 186 candidate intersection target genes of disease and MGNC were further screened to construct the interaction network. Gene ontology and KEGG pathway enrichment analysis ultimately unveiled a series of key targets that mainly mediated the effects of MGNC on DOR induced by chronic stress. The PI3K-Akt pathway may serve as the critical pathway underlying this therapeutic mechanism.
Conclusion: MGNC is a promising formula to treat DOR induced by chronic stress, and the PI3K-Akt pathway may play an essential role in this effect.

Keywords: modified Gengnianchun formula, diminished ovarian reserve, stress, follicle development, network pharmacology, target gene


中文翻译：

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基于实验方法和网络药理学的更年春方对应激性卵巢储备功能减退的治疗作用

目的：验证中药更年春方（MGNC）对卵巢储备减少（DOR）动物模型的影响，并通过网络药理学策略预测其潜在机制。
方法：性成熟雌性C57BL/6小鼠分为5组，简称对照组（C）组、应激模型组（M）、生理盐水灌胃应激（N）组、低剂量MGNC灌胃应激（L）组，高剂量 MGNC 灌胃 (H) 组应激。在应激和管饲过程中监测体重和发情周期。通过ELISA评估血清应激激素和生殖激素。计算卵巢卵泡计数，并通过蛋白质印迹和免疫组织化学评估卵巢卵泡刺激素受体 (FSHR) 和抗苗勒管激素 (AMH) 的表达。网络药理学策略包括活性化合物筛选、药物和疾病靶点分析、基因本体分析、通路分析和结果可视化。
结果：与 M 组和 N 组相比，MGNC 治疗显着降低了 H 组的血清皮质酮 (CORT) 和促卵泡激素 (FSH) 水平，并增加了睾酮 (T) 水平。与 M 组和 N 组相比，H 组的原始和窦前卵泡计数以及卵巢 AMH 和 FSHR 表达显着增加。通过药代动力学筛选，我们在 MGNC 中发现了 244 种活性化合物。进一步筛选出186个疾病与MGNC的候选交叉靶基因，构建交互网络。基因本体和KEGG通路富集分析最终揭示了一系列主要介导MGNC对慢性应激诱导的DOR影响的关键靶点。PI3K-Akt 通路可能是这种治疗机制的关键通路。
结论： MGNC是一种很有前景的治疗慢性应激诱导的DOR的配方，PI3K-Akt通路可能在该作用中发挥重要作用。

【关键词】：更年春方 卵巢储备功能减退 应激 卵泡发育 网络药理学 靶基因