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This study systematically characterized the chemical composition of Yuanhu Zhitong Oral Liquid (YZOL) and elucidated its therapeutic mechanism in primary dysmenorrhea (PD). Using UPLC-Q-TOF-MS/MS, a total of 100 constituents were preliminarily identified in YZOL, predominantly isoquinoline alkaloids and coumarins. A rat model of PD was established by combined administration of estradiol benzoate and oxytocin, and the pharmacological effects of YZOL were comprehensively evaluated through behavioral assessment, quantification of dysmenorrhea-related mediators, and histopathological examination. The results demonstrated that YZOL significantly reduced writhing frequency and prolonged writhing latency in PD rats (P < 0.05). Specifically, medium- and high-dose YZOL markedly decreased the PGF2α/PGE2 ratio (P < 0.05), while all dose groups significantly reduced the TXB2/6-keto-PGF1α ratio (P < 0.05). Consistently, YZOL treatment markedly ameliorated uterine histopathological damage, with the high-dose group exhibiting tissue morphology closest to normal. Transcriptomic profiling of uterine tissues identified 403 genes reversed by YZOL treatment, including 280 upregulated and 123 downregulated genes in the YZOL-H versus PD comparison. KEGG pathway enrichment analysis indicated that these reversed genes were primarily involved in the TGF-β, Rap1, PI3K-Akt, and MAPK signaling pathways, as well as ovarian steroidogenesis. Functional annotation further revealed that these pathways are closely associated with nociceptive signaling, inflammatory responses, synaptic activity, and hormone biosynthesis, and that their aberrant activation contributes to pain sensitization, inflammatory mediator release, and excessive uterine smooth muscle contraction during PD progression. Integrative analysis combining transcriptomics and network pharmacology identified seven core targets associated with key pathological processes, including pain transmission, inflammatory regulation, and uterine smooth muscle contraction. Molecular docking analysis using AutoDock Vina demonstrated that berberine, peucedanoside, bakuchiol alcohol, and other constituents exhibited strong binding affinities toward these targets, with binding energies comparable to or exceeding those of positive control compounds. In addition, several compounds showed favorable oral bioavailability, suggesting their potential contribution to the in vivo efficacy of YZOL. Based on bioinformatics analysis and batch molecular docking, a multidimensional "active component-key target-biological effect" network was constructed to systematically illustrate the therapeutic mechanism of YZOL against PD. Collectively, these findings indicate that YZOL exerts anti-dysmenorrheic effects through a multi-component, multi-target, and multi-pathway mode of action, in which key constituents including berberine, peucedanoside, bakuchiol alcohol, dihydrochelerythrine, and imperatorin may act on targets such as IL1B, PRKCG, TRPV1, and PTGER2 to coordinately modulate pain signaling, inflammatory responses, and uterine smooth muscle contraction. The experiment was approved by the Laboratory Animal Ethics Committee of the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences (approval No. 2021B020).
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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