背景:过早卵巢功能不全[POI]是一种以40岁之前卵巢功能过早下降为特征的疾病。在中国,女贞子[FLL]长期用于改善卵巢功能和治疗POI。
方法:本研究旨在通过网络药理学验证FLL对POI的影响,分子对接,和体外细胞实验。
结果:在FLL中筛选了13种活性物质,包括槲皮素,taxifolin,木犀草素,山奈酚,和β-谷甾醇.然后,网络分析发现,FLL可能通过10个靶标对POI产生影响,包括AR,ESR1,ESR2,KDR,CYP19A1,CLPP,GC,MMP3,PPARG,STS。根据GO和KEGG富集分析,FLL与雌激素相关的机制有关,包括类固醇激素的生物合成,卵巢类固醇生成,和雌激素信号通路。分子对接证实了FLL活性成分与编码芳香酶的CYP19A1之间的相互作用。CCK8实验证实槲皮素和紫杉醇林能增进KGN颗粒细胞的增殖,而槲皮素,taxifolin,山奈酚可抑制KGN颗粒细胞凋亡。ELISA实验证实槲皮素,taxifolin,木犀草素,山奈酚可以增加KGN颗粒细胞中雌二醇的合成。WB证实槲皮素可以增进KGN细胞中CYP19A1的表达程度。
结论:FLL可以促进细胞增殖,凋亡,卵巢颗粒细胞中雌二醇的合成,并有可能治疗POI。
BACKGROUND: Premature ovarian insufficiency [POI] is a disease characterized by a premature decline in ovarian function before the age of 40. In China, Ligustrum lucidum [FLL] has long been used to improve ovarian function and treat POI.
METHODS: This study aims to verify the effect of FLL on POI through network pharmacology, molecular docking, and in-vitro cell experiments.
RESULTS: A total of 13 active substances were screened in FLL, including including quercetin, taxifolin, luteolin, kaempferol, and beta-sitosterol. Then, network analysis found that FLL may exert effects on POI through 10 targets, including AR, ESR1, ESR2, KDR, CYP19A1, CLPP, GC, MMP3, PPARG, and STS. According to
GO and KEGG enrichment analysis, FLL is associated with mechanisms related to estrogen, including steroid hormone biosynthesis, ovarian steroidogenesis, and the estrogen signaling pathway. Molecular docking confirms the interaction between the active ingredients of FLL and CYP19A1, which encodes aromatase. CCK8 experiment confirmed that quercetin and taxifolin can enhance the proliferation of KGN granulosa cells, while quercetin, taxifolin, and kaempferol can inhibit the apoptosis of KGN granulosa cells. ELISA experiments have confirmed that quercetin, taxifolin, luteolin, and kaempferol can increase the synthesis of estradiol in KGN granulosa cells. WB confirms that quercetin can increase the expression level of CYP19A1 in KGN cells.
CONCLUSIONS: FLL can improve the proliferation, apoptosis, and synthesis of estradiol in ovarian granulosa cells, and has the potential to treat POI.