调周滋阴方治疗早发性卵巢功能不全的疗效、安全性及作用机制

doi:10.12122/j.issn.1673-4254.2025.05.05

摘要/Abstract

摘要：

目的基于网络药理学探讨调周滋阴方治疗早发性卵巢功能不全（POI）的作用机制，通过动物实验进行验证，并设计临床对照试验验证调周滋阴方对该病的临床疗效。方法利用TCMSP、PubChem、Swiss Target Prediction、Genecards、OMIM等数据库筛选相应靶点，获取核心PPI，进行GO、KEGG富集分析，构建“疾病-通路-靶点-成分-药物”网络，采用AutoDock对核心活性成分和核心靶点进行分子对接验证。通过动物实验，采用Western blotting进一步验证网络药理学结果。设计临床对照试验，对照组予芬吗通治疗，试验组在此基础上加用调周滋阴方，通过观察两组内分泌指标（FSH、LH、E₂、AMH）、窦卵泡数、Kupperman评分、不良反应发生率及妊娠率，验证调周滋阴方治疗该病的临床有效性。结果通过网络药理学方法，得到调周滋阴方治疗早发性卵巢功能不全的潜在靶点有163个；核心活性成分主要为山奈酚、β-谷甾醇、木犀草素、槲皮素，核心靶点主要为SRC、TP53、STAT3、PIK3CA、MAPK3等；GO功能富集主要包括细胞运动的正调控、蛋白质磷酸化等；KEGG富集通路主要包括癌症通路、PI3K-Akt信号通路等；分子对接结果显示核心活性成分与核心靶点间均有良好的结合能力。利用动物实验对关键通路进行验证，结果显示，与POI模型组大鼠比较，调周滋阴方可升高POI大鼠卵巢组织中p-PI3K和p-Akt蛋白表达（P<0.05），验证了网络药理学的部分预测结果。临床试验显示，两组E₂较治疗前升高，FSH、LH较治疗前降低，Kupperman评分较治疗前降低（P<0.05），且试验组优于对照组（P<0.05）；两组窦卵泡数较治疗前均升高（P<0.05），试验组AMH较治疗前升高（P<0.05），试验组与对照组间的差异无统计学意义（P>0.05）；治疗结束4周后，试验组治疗效果优于同期对照组（P<0.05）。结论调周滋阴方具有多成分、多靶点、多途径的治疗特点，其中激活PI3K/Akt通路是其作用机制之一，从而改善POI患者卵巢储备功能，缓解临床症状，提高临床疗效，具有良好的临床应用前景。

关键词: 早发性卵巢功能不全, 调周滋阴方, 网络药理学, 实验验证, 临床观察

Abstract:

Objective To assess the efficacy and safety of Tiaozhou Ziyin (TZZY) recipe for treatment of premature ovarian insufficiency (POI) and explore the possible mechanisms. Methods We used bioinformatics analyses and network pharmacology to identify the main active ingredients in TZZY recipe and their core targets, which were verified by Western blotting. We tested the efficacy and safety of the recipe in 60 POI patients, who were randomized into control group (n=30) with Femoston treatment and TZZY group (n=30) with additional TZZY recipe treatment for 3 menstrual cycles. Results The core active ingredients of TZZY recipe included kaempferol, β-sitosterol, luteolin, and quercetin. The core targets included SRC, TP53, STAT3, PIK3CA, and MAPK3, which were involved in positive regulation of cell movement and protein phosphorylation, the cancer pathways and the PI3K-Akt signaling pathway. Molecular docking showed that the core active ingredients had good binding ability with the core targets. In female rat models of POI, TZZY recipe treatment significantly up-regulated ovarian expressions of p-PI3K and p-Akt proteins. In the clinical trial, treatment with Femoston and Femoston plus TZZY recipe both significantly increased E₂ levels and reduced FSH and LH levels and Kupperman scores of the patients, and the combined treatment produced significantly stronger effects. Both treatments increased the number of antral follicles of the patients, but the combined treatment also significantly increased the levels of AMH. Conclusion The therapeutic mechanism of TZZY recipe for POI involves multiple active ingredients, multiple therapeutic targets and multiple pathways, and activating the PI3K /Akt pathway is one of its main mechanisms of action, to improve ovarian reserve function, alleviate clinical symptoms, and enhance clinical efficacy in POI patients.

Key words: Premature ovarian insufficiency, Tiaozhou Ziyin recipe, Network pharmacology, Experimental verification, clinical observation

唐培培, 谈勇, 殷燕云, 聂晓伟, 黄菁宇, 左文婷, 李玉玲. 调周滋阴方治疗早发性卵巢功能不全的疗效、安全性及作用机制[J]. 南方医科大学学报, 2025, 45(5): 929-941.

Peipei TANG, Yong TAN, Yanyun YIN, Xiaowei NIE, Jingyu HUANG, Wenting ZUO, Yuling LI. Tiaozhou Ziyin recipe for treatment of premature ovarian insufficiency: efficacy, safety and mechanism[J]. Journal of Southern Medical University, 2025, 45(5): 929-941.

图/表 17

参考文献 27

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TCM		MOL ID	Molecule name	OB (%)	DL
Shudi baishao shanzhuyu	A1	MOL000359	Sitosterol	36.91	0.75
Shudi shanzhuyu sharen	B1	MOL000449	Stigmasterol	43.83	0.76
Baishao shanzhuyu tusizi sharen	C1	MOL000358	Beta-sitosterol	36.91	0.75
Baishao tusizi	D1	MOL000422	Kaempferol	41.88	0.24
Shanzhuyu danshen sharen	E1	MOL001771	Poriferast-5-en-3beta-ol	36.91	0.75
Baishao	BS1	MOL001919	(3S,5R,8R,9R,10S,14S)-3,17-dihydroxy-4,4,8,10,14-pentamethyl-2,3,5, 6,7,9-hexahydro-1H-cyclopenta[a]phenanthrene-15,16-dione	43.56	0.53
Baishao	BS2	MOL001924	Paeoniflorin	53.87	0.79
Shanzhuyu	SZY1	MOL001494	Mandenol	42	0.19
Shanzhuyu	SZY2	MOL001495	Ethyl linolenate	46.1	0.2
Tusizi	TSZ1	MOL001558	Sesamin	56.55	0.83
Tusizi	TSZ8	MOL000098	Quercetin	46.43	0.28
Danshen	DS1	MOL001601	1,2,5,6-Tetrahydrotanshinone	38.75	0.36
Danshen	DS8	MOL000006	Luteolin	36.16	0.25
Sharen	SR1	MOL001755	24-Ethylcholest-4-en-3-one	36.08	0.76
Sharen	SR2	MOL001973	Sitosteryl acetate	40.39	0.85

TCM		MOL ID	Molecule name	OB (%)	DL
Shudi baishao shanzhuyu	A1	MOL000359	Sitosterol	36.91	0.75
Shudi shanzhuyu sharen	B1	MOL000449	Stigmasterol	43.83	0.76
Baishao shanzhuyu tusizi sharen	C1	MOL000358	Beta-sitosterol	36.91	0.75
Baishao tusizi	D1	MOL000422	Kaempferol	41.88	0.24
Shanzhuyu danshen sharen	E1	MOL001771	Poriferast-5-en-3beta-ol	36.91	0.75
Baishao	BS1	MOL001919	(3S,5R,8R,9R,10S,14S)-3,17-dihydroxy-4,4,8,10,14-pentamethyl-2,3,5, 6,7,9-hexahydro-1H-cyclopenta[a]phenanthrene-15,16-dione	43.56	0.53
Baishao	BS2	MOL001924	Paeoniflorin	53.87	0.79
Shanzhuyu	SZY1	MOL001494	Mandenol	42	0.19
Shanzhuyu	SZY2	MOL001495	Ethyl linolenate	46.1	0.2
Tusizi	TSZ1	MOL001558	Sesamin	56.55	0.83
Tusizi	TSZ8	MOL000098	Quercetin	46.43	0.28
Danshen	DS1	MOL001601	1,2,5,6-Tetrahydrotanshinone	38.75	0.36
Danshen	DS8	MOL000006	Luteolin	36.16	0.25
Sharen	SR1	MOL001755	24-Ethylcholest-4-en-3-one	36.08	0.76
Sharen	SR2	MOL001973	Sitosteryl acetate	40.39	0.85

Sub group	GO term	-LogP
BP	Positive regulation of locomotion	57.01529
BP	Positive regulation of cell motility	56.17373
BP	Positive regulation of cell migration	55.77701
BP	Protein phosphorylation	55.0968
BP	Positive regulation of phosphorylation	54.99759
BP	Response to hormone	54.48073
BP	Transmembrane receptor protein tyrosine kinase signaling pathway	47.79846
BP	Positive regulation of protein phosphorylation	47.32929
BP	Response to peptide	47.32309
BP	Enzyme-linked receptor protein signaling pathway	46.18651
CC	Membrane raft	22.13493
CC	Membrane microdomain	22.10097
CC	Receptor complex	17.86078
CC	Caveola	15.1191
CC	Focal adhesion	13.80176
CC	Cell-substrate junction	13.60269
CC	Plasma membrane raft	13.26789
CC	Side of membrane	12.02636
CC	External side of plasma membrane	10.71497
CC	Extracellular matrix	10.53049
MF	Protein kinase activity	53.06992
MF	Phosphotransferase activity, alcohol group as acceptor	48.91392
MF	Kinase activity	48.35798
MF	Protein tyrosine kinase activity	35.63223
MF	Kinase binding	28.34491
MF	Transmembrane receptor protein tyrosine kinase activity	28.34218
MF	Transmembrane receptor protein kinase activity	27.83784
MF	Protein serine/threonine kinase activity	25.19876
MF	Protein serine kinase activity	23.51593
MF	Protein kinase binding	22.3736

Sub group	GO term	-LogP
BP	Positive regulation of locomotion	57.01529
BP	Positive regulation of cell motility	56.17373
BP	Positive regulation of cell migration	55.77701
BP	Protein phosphorylation	55.0968
BP	Positive regulation of phosphorylation	54.99759
BP	Response to hormone	54.48073
BP	Transmembrane receptor protein tyrosine kinase signaling pathway	47.79846
BP	Positive regulation of protein phosphorylation	47.32929
BP	Response to peptide	47.32309
BP	Enzyme-linked receptor protein signaling pathway	46.18651
CC	Membrane raft	22.13493
CC	Membrane microdomain	22.10097
CC	Receptor complex	17.86078
CC	Caveola	15.1191
CC	Focal adhesion	13.80176
CC	Cell-substrate junction	13.60269
CC	Plasma membrane raft	13.26789
CC	Side of membrane	12.02636
CC	External side of plasma membrane	10.71497
CC	Extracellular matrix	10.53049
MF	Protein kinase activity	53.06992
MF	Phosphotransferase activity, alcohol group as acceptor	48.91392
MF	Kinase activity	48.35798
MF	Protein tyrosine kinase activity	35.63223
MF	Kinase binding	28.34491
MF	Transmembrane receptor protein tyrosine kinase activity	28.34218
MF	Transmembrane receptor protein kinase activity	27.83784
MF	Protein serine/threonine kinase activity	25.19876
MF	Protein serine kinase activity	23.51593
MF	Protein kinase binding	22.3736

ID	Pathway	GeneRatio	P	Count
hsa05200	Pathways in cancer	0.453988	6.19E-86	74
hsa04151	PI3K-Akt signaling pathway	0.300613	2.97E-55	49
hsa05205	Proteoglycans in cancer	0.245399	3.2E-51	40
hsa05215	Prostate cancer	0.196319	3.38E-49	32
hsa01521	EGFR tyrosine kinase inhibitor resistance	0.171779	3.99E-44	28
hsa05417	Lipid and atherosclerosis	0.214724	8.47E-42	35
hsa01522	Endocrine resistance	0.171779	5.01E-41	28
hsa04933	AGE-RAGE signaling pathway in diabetic complications	0.171779	9.62E-41	28
hsa05206	MicroRNAs in cancer	0.233129	1.66E-40	38
hsa04015	Rap1 signaling pathway	0.202454	7.19E-39	33
hsa04014	Ras signaling pathway	0.208589	1.07E-38	34
hsa04010	MAPK signaling pathway	0.220859	2.44E-38	36
hsa05163	Human cytomegalovirus infection	0.196319	3.1E-36	32
hsa05230	Central carbon metabolism in cancer	0.141104	1.92E-35	23
hsa05218	Melanoma	0.141104	4.14E-35	23
hsa04218	Cellular senescence	0.171779	9.59E-35	28
hsa05161	Hepatitis B	0.171779	2.98E-34	28
hsa05207	Chemical carcinogenesis - receptor activation	0.184049	6.44E-34	30
hsa05167	Kaposi sarcoma-associated herpesvirus infection	0.177914	1.6E-33	29
hsa05223	Non-small cell lung cancer	0.134969	4.06E-33	22