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Integrated metabolomics and network pharmacology to reveal the therapeutic mechanism of Dingkun Pill on polycystic ovary syndrome

發(fā)布時間:1668145780 人氣: 來源:

期刊名:Journal of Ethnopharmacology

發(fā)表日期:15 September 2022

文章地址https://doi.org/10.1016/j.jep.2022.115442


ABSTRACT

Ethnopharmacological relevance: Dingkun Pill (DKP), a traditional Chinese medicine prescription, was modifiedfrom Bujing decoction and Xusijiangsheng pill by the imperial physician in the Qing dynasty (1700 ′ s). It wasbelieved to treat various gynecological diseases by nourishing qi and blood. Accumulating evidence indicates that it is effective in treating polycystic ovary syndrome (PCOS). However, the therapeutic efficacy and mech-anism of action DKP against PCOS need to be further elucidated.


Aim of the study: To investigate the therapeutic effect and action mechanism of DKP against PCOS using an in-tegrated approach of metabolomics and network pharmacology.


Materials and methods: The rat model of PCOS was established by dehydroepiandrosterone. An integratedmetabolomics and network pharmacology strategy was applied to ystemically clarify the mechanism of DKP against PCOS. Theca cells were prepared to evaluate the effect of DKP and its ingredients on testosterone syn-thesis in vitro.


Results: The pharmacological experiments demonstrated that DKP could effectively convert the disorderedestrous cyclicity, decrease the level of testosterone and the luteinizing hormone/follicle stimulating hormoneratio, and inhibit abnormal follicle formation in PCOS rats. By metabolomics analysis, 164 serum endogenous differential metabolites and 172 urine endogenous differential metabolites were tentatively identified. Steroidhormone biosynthesis and ovarian steroidogenesis were the most significantly impacted pathways. Based onnetwork pharmacology and metabolomics studies, the ingredient-target-pathway network of DKP in the treat-ment of PCOS was constructed. Among the 10 key targets, CYP17A1, CYP19A1, STS, AR, ESR1, and MYC were closely involved in ovarian androgen synthesis. In theca cell-based assay of testosterone synthesis, DKP and its two active compounds (ligustilide and picrocrocin) showed inhibitory effects.


Conclusion: DKP effectively improved symptoms in rats with dehydroepiandrosterone-induced PCOS. The mechanism of DKP in the treatment of PCOS is related to the CYP17A1 enzyme required for androgen synthesis.


2. Materials and methods


2.1. Chemicals and reagents


The components of DKP (Shanxi GuangYuYuan Chinese Herbal Medicine Co., Ltd. Batch Number: Q20190282) are presented in Table 1.LC-MS purity grade formic acid, acetonitrile, and methanol were ac-quired from Fisher Scientific (Fair Lawn, NJ, USA). LC-MS-grade leucine enkephalin was obtained from Waters (Milford, USA). Reference stan-dards of corydaline, liquiritigenin, ligustilide, and isoliquiritigenin were obtained from Chengdu Pufei De Biotech Co., Ltd (Chengdu, China).Baohuoside I (internal standard 1, IS1), cavidine (IS2), baicalin, ferulic acid

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