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. 2025 Jul 2:16:1591558.
doi: 10.3389/fphar.2025.1591558. eCollection 2025.

Interaction of blood-entry components, network pharmacology and transcriptomics to elucidate the mechanism of Wentong plaster in treating primary dysmenorrhea

Zongtong Yang  1 , Ziqi Jiao  1 , Cheng Wang  1 , Xiaojing Li  1 , Mengyu Yuan  2 , Zaiyun Sui  1 , Wenhui Wang  3 , Wenjing Hou  1
Affiliations

Interaction of blood-entry components, network pharmacology and transcriptomics to elucidate the mechanism of Wentong plaster in treating primary dysmenorrhea

Zongtong Yang et al. Front Pharmacol. .

Abstract

Introduction: Primary dysmenorrhea (PD) is characterized by pain during the menstrual cycle, affects women's health. Our group developed a traditional Chinese medicine plaster (Wentong plaster, WTT) for the treatment of PD. However, the underlying mechanisms have not yet been elucidated.

Methods: In this study, the blood-entry components of WTT were detected using UPLC-Q-Exactive Orbitrap-MS, and the therapeutic functions of WTT on PD were evaluated by the writhing response, pathological analysis, and the levels of estrogen, nitric oxide, progesterone, among other indicators. Network pharmacology and transcriptomics were used to elucidate the underlying mechanisms. Finally, enzyme-linked immunosorbent assay and western blotting were used to determine the levels of relevant indicators.

Results: Our findings indicate that 49 original blood-entry components were detected. Meanwhile, WTT upregulated the level of NO, and downregulated the levels of PGF2α, PGE2, estrogen, and progesterone, thereby increasing blood flow, alleviating inflammatory responses, and inhibiting the writhing response. Results from network pharmacology and transcriptomics analyses indicated that WTT could increase the expression of Lcn2 and decrease the expression of Cxcl6 and IL-17, thereby regulating the IL-17 signaling pathway, and alleviating inflammation to treat PD.

Conclusion: WTT mainly down-regulates the levels of Cxcl6 and IL-17 and up-regulates the expression of Lcn2, further regulates the IL-17 signaling pathway to alleviate inflammation, ultimately treating PD. This study provides a basis for further research on the mechanism of WTT, and offers a reference for its clinical application.

Keywords: Wentong plaster; mechanism; network pharmacology; primary dysmenorrhea; transcriptomics.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Blood flow in uterine (A), claw (B), and sublingual (C). Compared with CG group: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; compared with PD group: #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001. CG, control group; PD, primary dysmenorrhea model group; WJZT, Wenjing Zhitong plaster treatment group; WTT, Wentong plaster treatment group.
FIGURE 2
FIGURE 2
Effect of WTT on levels of PGF2α (A), PGE2 (B), PGF2α/PGE2 relative expression (C), E2 (D), NO (E), and PG (F). Compared with CG group: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; compared with PD group: #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001. CG, control group; PD, primary dysmenorrhea model group; WJZT, Wenjing Zhitong plaster treatment group; WTT, Wentong plaster treatment group; E2, estrogen; PG, progesterone.
FIGURE 3
FIGURE 3
Histopathological analysis of rats. CG, control group, 10× (A), 40× (B); PD, primary dysmenorrhea model group, 10× (C), 40× (D); WJZT, Wenjing Zhitong plaster treatment group, 10× (E), 40× (F); WTT, Wentong plaster treatment group, 10× (G), 40× (H).
FIGURE 4
FIGURE 4
Effects of WTT on PT (A), APTT (B), TT (C), and FIB (D). Compared with CG group: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; compared with PD group: #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001. CG, control group; PD, primary dysmenorrhea model group; WJZT, Wenjing Zhitong plaster treatment group; WTT, Wentong plaster treatment group; PT, prothrombin time; FIB, fibrinogen, APTT, activated partial thromboplastin time; TT, thrombin time.
FIGURE 5
FIGURE 5
Network pharmacology analysis: Venny plot (A); PPI network (B); GO enrichment analysis (C); KEGG enrichment analysis (D); “Components Targets-Pathways” network (E).
FIGURE 6
FIGURE 6
The volcano plot of DEGs screened between PD and CG groups (A) and between WTT and PD groups (B); Venny plot (C); The KEGG enrichment analysis of DEGs screened between PD and CG groups (D) and between WTT and PD groups (E).
FIGURE 7
FIGURE 7
The levels of Cxcl6 (A), Ccl20 (B), Lcn2 (C), and IL-17 (D) in uterine tissue of PD rats. Compared with CG group: *: p < 0.05, **: p < 0.01; ***: p < 0.001; ****: p < 0.0001; Compared with PD group: #: p < 0.05, ##: p < 0.01; ###: p < 0.001; ####: p < 0.0001.
FIGURE 8
FIGURE 8
Effect of WTT on Cxcl6 and Lcn2 protein levels in PD rats determined by WB (A), and the ratio of Cxcl6 (B) and Lcn2 (C) to β-actin was calculated. Compared with CG group: *: p < 0.05, **: p < 0.01; ***: p < 0.001; ****: p < 0.0001; Compared with PD group: #: p < 0.05, ##: p < 0.01; ###: p < 0.001; ####: p < 0.0001.
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