Donghua Li

Curdione inhibits the proliferation of human uterine leiomyomas by targeting YTHDF1

Curdione, an active sesquiterpene found in Curcumae Rhizoma, exhibits strong anti-inflammatory and antitumor properties, but its mechanism of action against uterine leiomyomas (ULMs) is not yet clear. This study aims to clarify the anti-proliferative effects of curdione on ULMs and explore the molecular mechanisms, with specific focus on the role of the m6A reader protein YTHDF1. To evaluate the effects of curdione on human ULM cells, assays such as CCK-8, crystal violet staining, EdU, TUNEL, and Annexin V-FITC/PI were used to assess cell proliferation and apoptosis. Key m6A targets were identified through a comprehensive analysis involving GEO data integration, molecular docking, and surface plasmon resonance (SPR). RNA-seq and MeRIP-seq characterized N6-methyladenosine (m6A) methylation patterns and identify downstream regulatory targets. Lentiviral transfection was used to develop YTHDF1 knockdown and overexpression cell models, with target functions validated through molecular assays. The efficacy of curdione was assessed in a rat ULM model using histopathological analysis, electron microscopy, and molecular assays. Curdione exhibited an inhibitory effect on the viability of ULM cells, significantly reducing proliferation rates and inducing apoptosis. It was observed to downregulate YTHDF1 expression at both transcriptional and translational levels. MeRIP-seq analysis identified reprogramming of m6A methylation, with 231 hypermethylated and 984 hypomethylated peaks, alongside a reduction in THBS1 m6A modification and expression. Stable YTHDF1 knockdown and overexpression models were established through lentiviral transfection, confirming YTHDF1 as a pivotal mediator of curdione's anti-proliferative effects. In a rat ULM model, curdione administration resulted in a decreased uterine index, improved histopathological features, reduced collagen deposition, and normalization of serum inflammatory cytokines and sex hormones. Immunofluorescence and western blot analyses verified the co-localization and coordinated downregulation of YTHDF1 and THBS1 in uterine tissue. Curdione exerts its effects by downregulating YTHDF1, thereby influencing m6A modification and the translation of THBS1 mRNA. This study highlights curdione's potential as a therapeutic agent for ULMs and suggests that targeting YTHDF1 could be an effective management strategy.

Lichong decoction improves inflammatory microenvironment and alleviates fibrosis in uterine leiomyoma via targeting CXCL8

Lichong decoction (LD) is extensively employed in the treatment of uterine leiomyoma (ULM), demonstrating remarkable clinical effectiveness with an absence of notable adverse reactions. Its composition aligns with the traditional Chinese medicine (TCM) etiology of ULM, making it a highly suitable therapy. Nonetheless, the precise mechanisms underlying its therapeutic actions remain to be fully elucidated. The objective of this study was to clarify the therapeutic mechanism of LD improving ULM. The effects of LD on ULM cell viability, proliferation, and apoptosis were assessed using CCK-8, crystal violet staining, EdU incorporation, TUNEL, and Annexin V-FITC/PI assays. Gene microarray was used to profile differential gene expression after LD treatment. A rat ULM model was created to evaluate LD's anti-tumor efficacy, measuring body weight, uterine weight index, and sex hormone levels. Histopathological changes were analyzed with hematoxylin and eosin staining, Masson's trichrome staining, and transmission electron microscopy. Protein and RNA expression changes were analyzed via immunohistochemistry, western blotting, and qPCR. UHPLC-QE-MS enabled a detailed non-targeted LD analysis. Key components were identified through their correlation with serum sex hormones and inflammatory cytokines, and then examined by molecular docking studies. Experiments showed that LD reduced ULM cell viability and induced apoptosis. Gene expression profiling identified 313 differentially expressed genes. Enrichment analysis combined with experimental validation demonstrated that LD can reduce ULM fibrosis and inflammation by inhibiting the CXCL8/PI3K/AKT pathway. The analysis identified 494 primary compounds and 87 serum components in LD. Key compounds such as formononetin, palmatine, curcumenol, and hecogenin, which exhibit high affinity for CXCL8, may contribute to the anti-inflammatory and anti-tumor properties of LD. This study demonstrates that LD effectively inhibits ULM proliferation and fibrosis by improving the inflammatory microenvironment, primarily through the inhibition of CXCL8. These findings highlight the therapeutic potential of LD for ULM and provide new insights into its mechanisms.