Ju Shan, He Jialin, Wang Hanbi, Yang Liya, Guo AiXin, Guo Yiming, Qi Mingkang, Wang Huiping, Ai Lianzhong
School of Health Science and Engineering, Shanghai Engineering Research Center of Food Microbiology, University of Shanghai for Science and Technology, Shanghai, 200093, PR China.
Graduate School of Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, 100730, PR China; NHC Key Laboratory of Reproductive Health Engineering Technology Research(NRIFP), National Research Institute for Family Planning, Beijing, 100081, PR China.
J Ethnopharmacol. 2023 Mar 25;304:116054. doi: 10.1016/j.jep.2022.116054. Epub 2022 Dec 13.
The incidence of premature ovarian insufficiency (POI) is gradually increasing, the proportion is rising especially in female infertility patients. The risk of death of POI patients with cardiovascular disease also increases significantly. The cause of POI is complex and unclear, and clinical treatment is still in the exploratory stage, are two major constraints of treating POI. Traditional Chinese medicine (TCM) is widely used in the treatment of POI, and it is a good way to combine the development of modern new drugs with the help of TCM to predict the therapeutic targets.
In this study, four herbs commonly used in clinical treatment of POI, namely Radix Paeoniae, Polygonatum sibiricum, Rehmannia glutinosa and Eucommia ulmoides were selected to predict their mechanism in the treatment of POI, using network pharmacology methods. Then verify the predicted targets by animal test. Aim to find more effective POI potential core treatment targets and main pathways.
We screened the active ingredients of drugs from the TCM System Pharmacology Analysis Platform (TCMSP), Performed target prediction of active ingredients from databases such as SwissTargetPrediction and compare and analyze the POI-related targets retrieved from them to obtain potential targets for drug treatment of POI. Used STRING database to construct a protein interaction network, Cytoscape 3.7.2 software to construct an active ingredient-target-pathway network, and DAVID database to conduct the Kyoto Encyclopedia of Genes and Genomes (KEGG) on the intersection targets and gene ontology (GO) enrichment analysis.
The result is: there were 25 key targets for the treatment of POI with Radix Paeoniae Alba, 31 for the treatment of POI by Eucommia ulmoides, 28 for the treatment of POI by Polygonatum sibiricum, and 8 key targets for the treatment of Rehmannia glutinosa. The intersection targets of four herbs were defined as the core targets, which are CYP19A1, EGF, ESR1, ESR2, MDM2, AR, PCYP17A1, PPARG. Four Chinese herbs treat POI mainly through HIF-1 signaling pathway, PI3K-Akt signaling pathway, FoxO signaling pathway, Estrogen signaling pathway etc. A mouse model of POI was constructed based on the results of network pharmacology to verify the predicted targets. The results showed that the protein expression of the core target changed, and the estrogen level was increased by reducing the expression of peroxisome proliferator-activated receptor gamma (PPARG).
This study predicts the mechanism of multiple herbs in the treatment of POI, screens out more potential therapeutic drug targets and main pathways of POI treatment and provides new ideas for the subsequent development of POI therapeutic drugs.
卵巢早衰（POI）的发病率逐渐上升，在女性不孕症患者中的占比尤其升高。POI患者患心血管疾病的死亡风险也显著增加。POI的病因复杂且不明，临床治疗仍处于探索阶段，这是治疗POI的两大制约因素。中药在POI的治疗中被广泛应用，借助中药预测治疗靶点以开发现代新药是一种很好的方法。
本研究选用临床治疗POI常用的四味中药，即白芍、黄精、熟地黄和杜仲，采用网络药理学方法预测它们治疗POI的作用机制。然后通过动物实验验证预测靶点。旨在寻找更有效的POI潜在核心治疗靶点和主要通路。
我们从中药系统药理学分析平台（TCMSP）筛选药物活性成分，利用SwissTargetPrediction等数据库对活性成分进行靶点预测，并对从中检索到的POI相关靶点进行比较分析，以获得POI药物治疗的潜在靶点。使用STRING数据库构建蛋白质相互作用网络，利用Cytoscape 3.7.2软件构建活性成分-靶点-通路网络，并使用DAVID数据库对交集靶点进行京都基因与基因组百科全书（KEGG）和基因本体（GO）富集分析。
结果显示：白芍治疗POI有25个关键靶点，杜仲治疗POI有31个，黄精治疗POI有28个，熟地黄治疗POI有8个关键靶点。将四味中药的交集靶点定义为核心靶点，即CYP19A1、EGF、ESR1、ESR2、MDM2、AR、PCYP17A1、PPARG。四味中药治疗POI主要通过HIF-1信号通路、PI3K-Akt信号通路、FoxO信号通路、雌激素信号通路等。基于网络药理学结果构建POI小鼠模型以验证预测靶点。结果表明，核心靶点的蛋白表达发生变化，通过降低过氧化物酶体增殖物激活受体γ（PPARG）的表达使雌激素水平升高。
本研究预测了多味中药治疗POI的机制，筛选出更多POI治疗的潜在药物靶点和主要通路，为POI治疗药物的后续研发提供了新思路。
