目的 探索铜死亡相关基因在类风湿关节炎（rheumatoid arthritis，RA）中的表达及其作用机制，并筛选通过靶向铜死亡基因治疗RA的潜在中药。方法 通过检索GEO数据库获得RA芯片数据，分析铜死亡基因在RA中表达水平；依据铜死亡基因表达情况构建高表达亚型和低表达亚型，并进行差异分析，筛选出与RA相关的铜死亡基因。按不同分组对基因表达矩阵进行免疫浸润分析，并分析免疫浸润细胞与铜死亡基因之间的相关性。利用Kobas在线工具对差异基因进行京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析。通过Herb数据库检索靶向铜死亡基因治疗RA的潜在中药及其有效成分，使用Autodock vina软件模拟药物与铜死亡靶蛋白结合活性。通过收集临床样本验证铜死亡相关基因在RA中表达水平。结果 FDX1、DLD、DLAT、LIAS、GLS、LIPT1和PDHB 7个铜死亡相关基因在RA中差异表达。免疫浸润分析结果显示，在RA中活化的记忆CD4⁺ T细胞、静息自然杀伤（natural killer，NK）细胞和中性粒细胞与铜死亡相关。富集分析结果显示，铜死亡主要与趋化因子信号通路、Toll样受体信号通路、白细胞介素-17（interleukin-17，IL-17）信号通路、肿瘤坏死因子信号通路、核因子-κB信号通路、p53信号通路和过氧化物酶体增殖物激活受体信号通路相关。虎杖叶可能是靶向铜死亡基因治疗RA的潜在中药。分子对接结果显示，虎杖叶有效成分均有与铜死亡靶蛋白结合的可能。DLD、GLS、FDX1、DLAT在RA患者外周血单个核细胞中mRNA表达水平升高。结论 铜死亡相关基因表达水平与RA病程进展相关，并预测虎杖叶是潜在靶向铜死亡基因治疗RA的药物，为RA的临床诊疗及药物开发提供了新的方向和理论基础。

Objective To explore the expression of cuproptosis-related genes in rheumatoid arthritis (RA) and mechanism, and to screen the potential traditional Chinese medicine in the treatment RA by targeting cuproptosis-related genes. Methods The RA microarray data were obtained by searching the GEO database, and the expression levels of cuproptosis-related genes in RA were analyzed. The high expression subtypes and low expression subtypes were constructed according to the expression of cuproptosis-related genes, and differential analysis was performed to screen out the cuproptosis-related genes related to RA. Immune infiltration analysis was performed on the gene expression matrix by different groups and the correlation between immune infiltrating cells and cuproptosis-related genes was analyzed. The Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis was performed on differential expressed genes by the Kobas online tool. The Herb database was used to search for potential traditional Chinese medicines and their active ingredients in the treatment of RA by targeting cuproptosis-related gene, and the Autodock vina software was used to simulate the possible binding of drugs and cuproptosis-related target proteins. The expression levels of cuproptosis-related genes in RA were verified by clinical samples. Results A total of seven cuproptosis-related genes (FDX1, DLD, DLAT, LIAS, GLS, LIPT1, and PDHB) were differentially expressed in RA. The result of immune infiltration analysis showed that activated memory CD4⁺ T cells, resting NK cells, and neutrophils were associated with cuproptosis in RA. The results of pathway enrichment analysis showed that cuproptosis was mainly associated with the chemokine signaling pathway, Toll-like receptor signaling pathway, interleukin-17 (IL-17) signaling pathway, tumor necrosis factor signaling pathway, nuclear factor-κB signaling pathway, p53 signaling pathway, and peroxisome proliferators-activated receptors signaling pathways. Leaf of Huzhang (Polygonum cuspidatum) may be a potential drug in the treatment of RA by targeting cuproptosis-related gene, and the molecular docking results showed that the active components in leaf of P. cuspidatum may bind to cuproptosis-related proteins. The mRNA expression levels of DLD,GLS, FDX1 and DLAT in peripheral blood mononuclear cells of RA patients increased. Conclusion The expression level of cuproptosis -related genes is related to the progression of RA disease, and predicted that P. cuspidatum leaf is a potential drug in treatment of RA by targeting cuproptosis-related genes, which provides a new direction and theoretical basis for the clinical diagnosis and treatment of RA and drug development.

国家中医药管理局高水平中医药重点学科建设项目实施方案（国中医药人教函[2022]226号）；国家自然科学基金项目（82260880）；国家重点研发计划（2018YFC1705202，2018YFC1705203）；国家中医药管理局行业专项（20150700107）；甘肃省中医药项目（GZKP-2021-12）；兰州市科技指导性计划项目（2022-ZD-73）；王海东甘肃省名中医传承工作室建设项目（甘卫中医函[2022]50号）