Corydalis rhizoma attenuates primary dysmenorrhea in rats by inhibiting the FAK/PI3K-AKT/NF-κB signaling pathway

Abstract

Ethnopharmacological relevance: Primary dysmenorrhea (PD) is a prevalent condition affecting women globally. Corydalis rhizoma (known as Yanhusuo in Chinese, YHS) is commonly used to treat PD. However, the mechanisms by which raw-YHS (YHS-R) and vinegar-processed YHS (YHS-V) exert the therapeutic effects on PD remain unclear.

Aim of the study: To investigate the potential mechanisms of YHS-R and YHS-V in alleviating PD.

Materials and methods: UPLC-Q-TOF/MS technology was utilized to identify and quantify the components present in both YHS-R and YHS-V, as well as in the blood. The targets of these blood-absorbed components, in relation to PD, were subjected to KEGG enrichment analysis. A rat model of PD was established using estradiol benzoate and oxytocin. The therapeutic effects of YHS-R and YHS-V on PD were evaluated through the torsion response, uterine and ovarian indices, levels of PGE₂ and PGF_2α, and histopathological examination of the uterus and ovaries. Furthermore, the regulatory mechanisms of YHS-R and YHS-V on PD were investigated through RNA sequencing (RNA-seq), Western blotting, quantitative reverse transcription polymerase chain reaction (RT-qPCR), and non-targeted metabolomics.

Results: Both YHS-R and YHS-V significantly alleviated PD, as demonstrated by reduced writhing scores, improved uterine and ovarian indices, and decreased pathological damage to the uterus and ovaries. Additionally, they lowered the PGF_2α/PGE₂ ratio and inflammatory factor levels in the uterus and ovaries, with YHS-V showing a superior effect. Network pharmacology and RNA-seq analysis indicated that the PI3K-AKT and NF-κB pathways were involved in the PD-alleviating effects of YHS-R and YHS-V. Moreover, the glycerophospholipid and amino acid metabolism pathways were identified as significant metabolic pathways, the key metabolites PE and PG are closely correlated with the expression of the ITGB3, IL-24, and Thbs4 genes. Western blotting and RT-qPCR assays revealed that both YHS-R and YHS-V reduced the expression of FAK, PI3K, AKT, P65, and COX-2 in the uterus.

Conclusion: Collectively, both YHS-V and YHS-R alleviate PD through mechanisms involving suppression of inflammatory factor release, mitigation of uterine histopathological damage, and regulation of the FAK/PI3K-AKT/NF-κB signaling pathway and glycerophospholipid metabolism pathway. This study is the first to investigate the therapeutic effects of YHS-R and YHS-V in PD and elucidate the underlying mechanism, providing novel evidence supporting YHS application.

Keywords: Corydalis rhizoma; FAK/PI3K-AKT/NF-κB; Non-targeted metabolomics; Primary dysmenorrhea (PD); RNA-seq; Vinegar processing.