AUTHOR=Qin Yuwen , Zhang Wei , Bian Zhenhua , Fei Chenghao , Su Lianlin , Xue Rong , Zhang Qian , Li Yu , Chen Peng , Shi Yabo , Li Mingxuan , Mao Chunqin , Zhao Xiaoli , Ji De , Lu Tulin 

TITLE=The therapeutic mechanism of Curcumae Radix against primary dysmenorrea based on 5-HTR/Ca2+/MAPK and fatty acids metabolomics

JOURNAL=Frontiers in Pharmacology

VOLUME=14

YEAR=2023

URL=https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2023.1087654

DOI=10.3389/fphar.2023.1087654

ISSN=1663-9812

ABSTRACT=<p><bold>Background:</bold><italic>Curcumae</italic> Radix (CW) is traditionally used to treat primary dysmenorrea (PD). However, the mechanisms of action of CW in the treatment of PD have not yet been comprehensively resolved.</p><p><bold>Objective:</bold> To investigate the therapeutic effects of CW on PD and its possible mechanisms of action.</p><p><bold>Methods:</bold> An isolated uterine spastic contraction model induced by oxytocin was constructed in an <italic>in vitro</italic> pharmacodynamic assay. An animal model of PD induced by combined estradiol benzoate and adrenaline hydrochloride-assisted stimulation was established. After oral administration of CW, a histopathological examination was performed and biochemical factor levels were measured to evaluate the therapeutic effect of CW on PD. The chemical compositions of the drug-containing serum and its metabolites were analyzed by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry. Network pharmacology and serum untargeted metabolomics were used to predict the mechanism of CW treatment for PD, and the predicted results were validated by RT-qPCR, WB, and targeted fatty acid (FA) metabolism.</p><p><bold>Results:</bold><italic>In vitro,</italic> CW can relax an isolated uterus by reducing uterine motility. <italic>In vivo</italic>, the results showed that CW attenuated histopathological damage in the uterus and regulated PGF<sub>2α</sub>, PGE<sub>2</sub>, β-EP, 5-HT, and Ca<sup>2+</sup> levels in PD rats. A total of 66 compounds and their metabolites were identified in the drug-containing serum, and the metabolic pathways of these components mainly included hydrogenation and oxidation. Mechanistic studies showed that CW downregulated the expression of key genes in the 5-HTR/Ca<sup>2+</sup>/MAPK pathway, such as 5-HTR2A, IP3R, PKC, cALM, and ERK. Similarly, CW downregulated the expression of key proteins in the 5-HTR/Ca<sup>2+</sup>/MAPK pathway, such as p-ERK/ERK. Indirectly, it ameliorates the abnormal FA metabolism downstream of this signaling pathway in PD rats, especially the metabolism of arachidonic acid (AA).</p><p><bold>Conclusion:</bold> The development of PD may be associated with the inhibition of the 5-HTR/Ca<sup>2+</sup>/MAPK signaling pathway and FA metabolic pathways, providing a basis for the subsequent exploitation of CW.</p>