基于细胞异质性探索当归抗类风湿关节炎的作用机制

Exploration of the Mechanism of Danggui(Chinese angelica)against Rheumatoid Arthritis based on Cell Heterogeneity

目的:基于单细胞测序和分子对接技术探索当归抗类风湿关节炎的作用机制。方法:在GEO数据库获取类风湿关节炎单细胞外周血测序数据,并筛选出差异基因。通过对细胞进行PCA降维和UMAP聚类分析,同时进行细胞注释、寻找Marker基因和拟时序分析。筛选当归的有效成分,通过分子对接模拟药物与Marker基因在体内的相互作用。然后进一步体内实验验证,将15只Wistar大鼠随机分为正常组、模型组、当归组,每组5只。模型组、当归组大鼠构建类风湿关节炎模型,给药干预后观察各组大鼠关节组织病理学变化,利用RT-PCR和Western blotting检测差异基因的表达量。结果:差异基因有Ighm、Jchain、Elane等,主要分布在B细胞和红细胞中。类风湿关节炎的初始阶段涉及B细胞、CD8^(+)T细胞、单核细胞。当归的有效成分beta-sitosterol和Stigmasterol与类风湿关节炎Marker基因Ighm、Jchain具有较好的结合活性。当归组大鼠关节炎症明显低于模型组,且当归组大鼠Jchain mRNA和蛋白相对表达量均明显低于模型组(P<0.01或P<0.05)。结论:当归具有抗类风湿关节炎的作用,可能通过影响B细胞中Jchain的表达水平来调节免疫功能,对RA的临床防治具有重要意义。

Objective:To explore the anti-rheumatoid arthritis mechanism of Danggui(Chinese angelica)based on single cell sequencing and molecular docking technology.Methods:The single cell peripheral blood sequencing data of rheumatoid arthritis were obtained from GEO database,and the differential genes were screened.Through PCA dimension reduction and UMAP cluster analysis on cells,cell annotation,marker gene search and quasi timing analysis were conducted at the same time.The active components of Danggui were screened,and the interaction between drugs and Marker genes in vivo was simulated by molecular docking.Then conduct in vivo experimental verification,15 Wistar rats were randomly divided into normal group,model group and Danggui group,with 5 rats in each group.The model of rheumatoid arthritis was established in the model group and Danggui group.The histopathological changes of the joints in each group were observed after drug intervention,and the expression levels of differential genes were detected by RT-PCR and Western blotting.Results:The differentially expressed genes were Ighm,Jchain and Elane,which were mainly distributed in B cells and red blood cells.The initial stages of rheumatoid arthritis involve B cells,CD8^(+)T cells,and monocytes.Beta sitosterol and Stigmasterol,the active components of Danggui,had good binding activities with Ighm and Jchain genes of rheumatoid arthritis Marker.The joint inflammation of rats in Danggui group was significantly lower than that in model group,and the relative expression levels of Jchain mRNA and protein in Danggui group were significantly lower than those in model group(P<0.01 or P<0.05).Conclusion:Danggui(Chinese angelica)has the effect of anti-rheumatoid arthritis,which may regulate the immune function by affecting the expression level of Jchain in B cells,and has important significance for the clinical prevention and treatment of RA.