基于网络药理学及动物实验探究益肾祛浊方治疗PCOS-IR的作用机制

doi:10.12280/gjszjk.20250141

摘要/Abstract

摘要：

目的：通过网络药理学和实验验证，探讨益肾祛浊方治疗多囊卵巢综合征（polycystic ovary syndrome，PCOS）合并胰岛素抵抗（insulin resistance，IR）的作用机制。方法：检索中药系统药理学数据库与分析平台（TCMSP）数据库筛选益肾祛浊方的活性成分及作用靶点；利用GeneCards、OMIM、DrugBank等数据库筛选PCOS-IR疾病靶点；构建药物-成分-靶点网络；通过STRING和DAVID进行蛋白质-蛋白质互作（protein-protein interaction，PPI）网络分析、基因本体（Gene Ontology，GO）功能及京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）通路富集分析；使用AutodockTools 1.5.7软件进行分子对接。高脂饲料联合来曲唑构建PCOS-IR大鼠模型，检测血清性激素、糖代谢指标，并利用蛋白质印迹和实时荧光定量聚合酶链反应验证关键靶点表达。结果：检索到益肾祛浊方主要有效成分136个，靶点406个。选出山柰酚、槲皮素、β-谷甾醇等5种核心化学成分。核心网络中的靶点按照度值进行排名，筛选到白细胞介素-6（interleukin-6，IL-6）、信号转导及转录活化因子3（signal transducer and activator of transcription 3，STAT3）、蛋白激酶B1（protein kinase B1，AKT1）、热休克蛋白90（heat shock protein 90，HSP90）、肿瘤蛋白53（tumor protein 53，TP53）等10个关键靶点。富集于晚期糖基化终末产物-晚期糖基化终末产物受体（advanced glycation end product-receptor for advanced glycation end product，AGE-RAGE）、磷脂酰肌醇3激酶（phosphoinositide 3-kinase，PI3K）-AKT等信号通路。动物实验表明，益肾祛浊方可降低PCOS-IR大鼠血清黄体生成素（luteinizing hormone，LH）、睾酮、空腹胰岛素（fasting insulin，FINS）及稳态模型评估胰岛素抵抗指数（HOMA-IR）水平，差异均有统计学意义（均P<0.05），下调PCOS-IR大鼠卵巢组织STAT3、表皮生长因子受体（epidermal growth factor receptor，EGFR）、HSP90蛋白及基因表达，上调AKT1、TP53蛋白及基因表达，差异均有统计学意义（均P<0.05）。结论：益肾祛浊方通过多成分、多靶点、多通路改善PCOS-IR内分泌异常及糖代谢紊乱。

关键词: 多囊卵巢综合征, 胰岛素抵抗, 药理学, 分子对接模拟, 益肾祛浊方, 网络药理学

Abstract:

Objective: To investigate the mechanism of Yishen Quzhuo Formula in treating polycystic ovary syndrome with insulin resistance (PCOS-IR) through the network pharmacology and experimental validation. Methods: Active components and targets of Yishen Quzhuo Formula were retrieved from databases such as Traditional Chinese Medicine Systems Pharmacology (TCMSP). The targets of PCOS-IR-related disease were screened using GeneCards, OMIM, and DrugBank databases. A drug-component-target network was then constructed. Protein-protein interaction (PPI) network analysis, Gene Ontology (GO) function, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using STRING and DAVID. Molecular docking was conducted with AutodockTools 1.5.7 software. A PCOS-IR rat model was established using the high-fat diet combined with letrozole. Serum hormone levels and glucose metabolism indices were evaluated. The expression of key targets was validated via Western blotting and quantitative polymerase chain reaction (qPCR). Results: A total of 136 potential active components of Yishen Quzhuo Formula and 406 PCOS-IR-related targets were identified. Five core components, including kaempferol, quercetin, and β-sitosterol, were selected. The top 10 key targets, such as interleukin-6 (IL-6), signal transducer and activator of transcription 3 (STAT3), protein kinase B1 (AKT1), heat shock protein 90 (HSP90), and tumor protein 53 (TP53), were ranked by the degree values in the core network, enriched in the pathways like advanced glycation end product-receptor for advanced glycation end product (AGE-RAGE) and phosphoinositide 3-kinase (PI3K)-AKT. Animal experiments showed that Yishen Quzhuo Formula significantly reduced the levels of serum luteinizing hormone (LH), testosterone, fasting insulin (FINS), and homeostasis model assessment of insulin resistance (HOMA-IR) in PCOS-IR rats (all P<0.05), and downregulated the expressions of STAT3, epidermal growth factor receptor (EGFR) and HSP90 in the ovarian tissues of PCOS-IR rats, while upregulated the expressions of AKT1 and TP53 (all P<0.05). Conclusions: Yishen Quzhuo Formula ameliorates the endocrine abnormalities and glucose metabolism disorders of PCOS-IR through multi-component, multi-target and multi-pathway mechanisms.

Key words: Polycystic ovary syndrome, Insulin resistance, Pharmacology, Molecular docking simulation, Yishen Quzhuo formula, Network pharmacology

李梦元, 高征, 许昕. 基于网络药理学及动物实验探究益肾祛浊方治疗PCOS-IR的作用机制[J]. 国际生殖健康/计划生育杂志, 2025, 44(5): 353-360.

LI Meng-yuan, GAO Zheng, XU Xin. Mechanism of Yishen Quzhuo Formula in Treating PCOS-IR Based on Network Pharmacology and Animal Experiments[J]. Journal of International Reproductive Health/Family Planning, 2025, 44(5): 353-360.

图/表 13

参考文献 23

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基因	引物	序列（5′→3′）
GAPDH	正向	TCATTGACCTCAACTACATGG
	反向	TCGCTCCTGGAAGATGGTG
STAT3	正向	TTAACATTCTGGGCACGAACA
	反向	TGACAATCAAGGAGGCATCAC
AKT1	正向	CGACGTAGCCATTGTGAAGGAG
	反向	ATTGTGCCACTGAGAAGTTGTTG
TP53	正向	ATGGAGGATTCACAGTCGGATAT
	反向	CGCTGTGGTGGGCAGAATAT
EGFR	正向	CTCAGCAGGCCACACTTGTA
	反向	TGCATGGTTACTCCAGCTTAGA
HSP90	正向	ACATGCTGCGGAGGGTTAAG
	反向	CTGCTCGGAGTCATCTGTGGT

基因	引物	序列（5′→3′）
GAPDH	正向	TCATTGACCTCAACTACATGG
	反向	TCGCTCCTGGAAGATGGTG
STAT3	正向	TTAACATTCTGGGCACGAACA
	反向	TGACAATCAAGGAGGCATCAC
AKT1	正向	CGACGTAGCCATTGTGAAGGAG
	反向	ATTGTGCCACTGAGAAGTTGTTG
TP53	正向	ATGGAGGATTCACAGTCGGATAT
	反向	CGCTGTGGTGGGCAGAATAT
EGFR	正向	CTCAGCAGGCCACACTTGTA
	反向	TGCATGGTTACTCCAGCTTAGA
HSP90	正向	ACATGCTGCGGAGGGTTAAG
	反向	CTGCTCGGAGTCATCTGTGGT

编号	核心靶点	PDB ID	化合物	结合能（kcal/mol）
1	AKT1	8uw9	β-谷甾醇	-7.6
2	TP53	6vrn	山柰酚	-7.8
3	STAT3	6TLC	槲皮素	-8.6
4	EGFR	7zyn	7-甲氧基-2-甲基异黄酮	-6.4
5	HSP90	7s8y	木犀草素	-8.6

编号	核心靶点	PDB ID	化合物	结合能（kcal/mol）
1	AKT1	8uw9	β-谷甾醇	-7.6
2	TP53	6vrn	山柰酚	-7.8
3	STAT3	6TLC	槲皮素	-8.6
4	EGFR	7zyn	7-甲氧基-2-甲基异黄酮	-6.4
5	HSP90	7s8y	木犀草素	-8.6

组别	n	FSH （mIU/mL）	LH （mIU/mL）	LH/FSH 比值	睾酮（ng/mL）
空白组	8	1.766±0.317	7.438±0.374*	4.376±1.062*	0.643±0.032*
模型组	8	1.901±0.281	11.700±1.036	6.305±1.305	0.888±0.086
益肾祛浊方组	8	1.668±0.230	9.254±0.465*	5.656±0.918	0.763±0.086*
二甲双胍组	8	1.723±0.139	9.343±0.563*	5.456±0.582	0.758±0.068*
F		1.264	55.910	5.122	15.690
P		0.306	<0.001	0.006	<0.001