Department of TCM Gynecology, Hangzhou TCM Hospital of Zhejiang Chinese Medical University, Hangzhou, People's Republic of China.
School of Public Health, Zhejiang Chinese Medical University, Hangzhou, People's Republic of China.
Drug Des Devel Ther. 2023 Jul 24;17:2147-2163. doi: 10.2147/DDDT.S413142. eCollection 2023.
The aim of this study is to examine, using network pharmacology analysis and experimental validation, the pharmacological processes by which Yulin Formula (YLF) reduces cyclophosphamide-induced diminished ovarian reserve (DOR).
First, information about the active components, associated targets, and related genes of YLF and DOR was gathered from open-access databases. The primary targets and pathways of YLF to reduce DOR were predicted using studies of functional enrichment from the Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO), and Protein-Protein Interaction (PPI) networks. Second, we built a cyclophosphamide-induced diminished ovarian reserve (DOR) rat model to verify the primary target proteins implicated in the predicted signaling pathway to explore the mechanism of improve ovarian function of YLF.
98 targets met the targets of the 82 active ingredients in YLF and DOR after searching the intersection of the active ingredient targets and DOR targets. Fourteen targets, including AKT and Caspase-3 among others, were hub targets, according to the PPI network study. The PI3K/AKT pathway was revealed to be enriched by numerous targets by the GO and KEGG enrichment studies, and it was used as a target for in vivo validation. Animal studies showed that YLF administration not only reduced the number of atretic follicles, the proportion of TUNEL-positive ovarian cells, the rate of apoptosis of granulosa cells (GCs) and the proportion of abnormal mitochondria in DOR rats, but also reversed the high expression of Caspase-3, Caspase-9, BAX, cytochrome C, PI3K and P-AKT, improving the ovarian reserve in cyclophosphamide (CTX)-induced DOR rats.
Our research results predicted the active ingredients and potential targets of YLF-interfering DOR by an integrated network pharmacology approach, and experimentally validated some key target proteins participated in the predicted signaling pathway. A more comprehensive understanding of the pharmacological mechanism of YLF for DOR treatment was obtained.
本研究旨在通过网络药理学分析和实验验证，探讨玉林方（YLF）降低环磷酰胺诱导的卵巢储备减少（DOR）的药理过程。
首先，从公开数据库中收集 YLF 的活性成分、相关靶点和相关基因信息，以及 DOR 的信息。利用京都基因与基因组百科全书（KEGG）、基因本体论（GO）和蛋白质-蛋白质相互作用（PPI）网络的功能富集研究，预测 YLF 降低 DOR 的主要靶点和途径。其次，构建环磷酰胺诱导的卵巢储备减少（DOR）大鼠模型，验证预测信号通路中涉及的主要靶蛋白，探讨 YLF 改善卵巢功能的作用机制。
通过活性成分靶点和 DOR 靶点的交集搜索，98 个靶点符合 YLF 和 DOR 的靶点。根据 PPI 网络研究，14 个靶点，包括 AKT 和 Caspase-3 等，是枢纽靶点。GO 和 KEGG 富集研究表明，许多靶标富集了 PI3K/AKT 通路，该通路被用作体内验证的靶点。动物研究表明，YLF 给药不仅减少了 DOR 大鼠的闭锁卵泡数量、TUNEL 阳性卵巢细胞比例、颗粒细胞（GC）凋亡率和异常线粒体比例，而且逆转了 DOR 大鼠中 Caspase-3、Caspase-9、BAX、细胞色素 C、PI3K 和 P-AKT 的高表达，改善了环磷酰胺（CTX）诱导的 DOR 大鼠的卵巢储备。
本研究通过整合网络药理学方法预测了 YLF 干预 DOR 的活性成分和潜在靶点，并通过实验验证了一些参与预测信号通路的关键靶蛋白。更全面地了解了 YLF 治疗 DOR 的药理机制。
