类风湿关节炎（RAFLS）中的滑膜成纤维细胞表现出糖酵解和谷氨酰胺酶代谢的病理异常。因此，我们旨在研究植物生物化合物c28MS对这些途径的双重抑制是否具有通过靶向葡萄糖和谷氨酸代谢的潜力实现类风湿性关节炎的协同治疗。采用scRNA-seq评估了HK2和GLS在不同细胞类型和人类滑膜细胞以及小鼠关节炎模型中的相关基因表达。使用H1核磁共振波谱法在糖酵解和谷氨酸酶抑制条件下，通过与3-溴丙酮酸、CB839或双重抑制剂c28MS孵育，对RAFLS细胞进行代谢谱学分析。在类似条件下进行FLS功能分析。采用ELISA法测定IL-6、CCL2和MMP3的定量。将K/BxN血清注射到小鼠体内，引起关节炎进行体内类风湿性关节炎实验。scRNA-seq分析显示，许多成纤维细胞表达Hk2和Gls，并且一些基因包括Ptgs2、Hif1a、Timp1、Cxcl5和Plod2仅与双阳性成纤维细胞相关，表明双重抑制可能是成纤维细胞的一个具有吸引力的靶点。代谢组学和功能分析显示，c28MS通过靶向上调的糖酵解和谷氨酰胺酶代谢降低了RAFLS的侵袭性行为。在体内，c28MS明显减轻了K/BxN模型关节炎的严重程度。我们的发现表明，双重抑制糖酵解和谷氨酰胺酶可能是治疗RA的有效方法。它还暗示在非肿瘤性疾病中，靶向多个代谢途径可能是一种新颖的治疗方法。© 2023年。BioMed Central Ltd.是Springer Nature的一部分。

Synovial fibroblasts in rheumatoid arthritis (RAFLS) exhibit a pathological aberration of glycolysis and glutaminolysis. Henceforth, we aimed to investigate if dual inhibition of these pathways by phytobiological compound c28MS has the potential of synergistic therapy for arthritis by targeting both glucose and glutamine metabolism.The presence of HK2 and GLS across various cell types and associated gene expression in human synovial cells and a murine model of arthritis was evaluated by scRNA-seq. The metabolic profiling of RAFLS cells was done using H1-nuclear magnetic resonance spectroscopy under glycolytic and glutaminolytic inhibitory conditions by incubating with 3-bromopyruvate, CB839, or dual inhibitor c28MS. FLS functional analysis was conducted under similar conditions. ELISA was employed for the quantification of IL-6, CCL2, and MMP3. K/BxN sera was administered to mice to induce arthritis for in vivo arthritis experiments.scRNA-seq analysis revealed that many fibroblasts expressed Hk2 along with Gls with several genes including Ptgs2, Hif1a, Timp1, Cxcl5, and Plod2 only associated with double-positive fibroblasts, suggesting that dual inhibition can be an attractive target for fibroblasts. Metabolomic and functional analysis revealed that c28MS decreased the aggressive behavior of RAFLS by targeting both upregulated glycolysis and glutaminolysis. c28MS administered in vivo significantly decreased the severity of arthritis in the K/BxN model.Our findings imply that dual inhibition of glycolysis and glutaminolysis could be an effective approach for the treatment of RA. It also suggests that targeting more than one metabolic pathway can be a novel treatment approach in non-cancer diseases.© 2023. BioMed Central Ltd., part of Springer Nature.