Coniferyl aldehyde in ginger-Eucommiae Cortex enhances osteoarthritis treatment by modulating ALDOA and H3K23la...

Eucommiae Cortex (EC), a traditional Chinese medicinal herb, has been utilized to treat osteoarthritis (OA). The therapeutic efficacy of EC can be augmented by combining it with ginger juice. OA is often linked to metabolic disorders, with lactate accumulation contributing to its progression. Notably, the concentration of coniferyl aldehyde (CFA) markedly increases during the processing of EC with ginger juice. This study seeks to investigate whether CFA ameliorates OA via lactylation and to develop a non-invasive machine-learning model for the rapid diagnosis of OA utilizing lactate and other metabolic markers.

Differences in the chemical composition of EC before and after processing with ginger were investigated using an untargeted metabolomics strategy and database integration. Active components were screened using network pharmacology to examine CFA's effect on inflammation in tumor necrosis factor-α (TNF-α) -induced C28/I2 and primary mouse chondrocytes. Biomarkers and molecular mechanisms were identified using metabolomics, interpretable machine learning, and RNA sequencing. In-cell western blotting and expression correlation analysis confirmed the correlation between inflammation and H3 histone lactylation improvement in C28/I2 cells treated with CFA. Molecular docking analysis identified targets, and the overexpression (oe) and silencing (si) of ALDOA validated the inhibitory effect of CFA on lactylation. The in vivo efficacy of CFA was assessed using a mouse destabilization of the medial meniscus (DMM) model of OA.

CFA, a principal component in G-EC, exhibited significant anti-inflammatory effects both in vitro and in vivo. A non-invasive diagnostic model utilizing machine learning was developed by analyzing 44 OA urine samples from the Metabolomics Workbench database. This model employed SHapley Additive exPlanations in conjunction with a random forest algorithm and identified lactate as a significant potential diagnostic metabolite for OA. Metabolomic analysis of C28/I2 cells indicated that CFA influences the glycolysis pathway.

This study identified the basis for the synergistic effect of ginger juice on EC and supported the scientific rationale for G-EC processing. In this study, CFA improved OA by affecting glycolysis and modulating lactylation at the H3K23la site of H3 histones, highlighting the critical role of CFA in its anti-OA effects. A non-invasive diagnostic model for OA was developed, which facilitated the rapid prediction of OA risk in patients, and interpretable machine-learning methodologies enabled the analysis of key metabolic markers.