UNASSIGNED: This study adopted the method of network pharmacology to filter key gene targets of Juanbi recipe in RA treatment. Single-cell ribonucleic acid (RNA) sequencing data was used to screen the key genes to form the core genes of Juanbi recipe in RA treatment. The molecular docking technique was used to verify the core target genes and explore the mechanisms of Juanbi recipe in RA treatment. The RA model of mice was induced by the collagen-induced arthritis and the effect of Juanbi recipe was evaluated by intragastric administrating of extraction of Juanbi recipe. Enzyme linked immunosorbent assay was used to analysis serum inflammatory factors. Hematoxylin and eosin staining was used to evaluate inflammation and immunohistochemical (IHC) staining was used to evaluate core target genes and pathways in synovium of ankle.
UNASSIGNED: This study screened out 281 active molecules in Juanbi recipe, found 105 key target genes of Juanbi recipe in RA treatment, and drew an \"ingredient - molecule - gene\" diagram. Juanbi recipe reduced the levels of serum interleukin (IL)-1 and IL-6, the inflammatory infiltration in synovium, demonstration that Juanbi recipe reduced both systemic and synovial inflammatory response. Single cell RNA sequencing data were used to select six core target genes and six core active molecules of Juanbi recipe in RA treatment. The pathways of Juanbi recipe in RA treatment involved in activator protein-1 (AP-1) and nuclear factor kappa B (NF-κB) pathway. Results of western blot and IHC staining showed that Juanbi recipe decreased the expressions of c-jun and p65, which demonstrated that Juanbi recipe inhibited the expression of AP-1 and NF-κB pathway in RA.
UNASSIGNED: The core active molecules of Juanbi recipe could inhibit key factors of AP-1 and NF-κB pathway to inhibit the inflammation, which played a protective role in RA.
■本研究采用网络药理学的方法,筛选了在类风湿关节炎治疗中的关键基因靶点。利用单细胞核糖核酸(RNA)测序数据筛选出关键基因,形成Juanbi方治疗RA的核心基因。采用分子对接技术对核心靶基因进行验证,并初步探讨Juanbis方治疗RA的作用机制。采用胶原诱导的关节炎模型建立小鼠RA模型,并通过灌胃提取的juanbis方评价juanbis方的效果。采用酶联免疫吸附试验分析血清炎症因子。苏木精和伊红染色用于评估炎症,免疫组织化学(IHC)染色用于评估踝关节滑膜中的核心靶基因和通路。
■这项研究筛选出了Juanbi配方中的281个活性分子,在RA治疗中发现了Juanbi方的105个关键靶基因,并绘制了一张“成分-分子-基因”图。Juanbi方降低血清白细胞介素(IL)-1和IL-6的水平,滑膜中的炎症浸润,证明Juanbi方降低了全身和滑膜炎症反应。单细胞RNA测序数据用于选择RA治疗中的6个核心靶基因和6个核心活性分子。Juanbi方治疗RA的途径涉及激活蛋白-1(AP-1)和核因子κB(NF-κB)途径。westernblot和IHC染色结果表明,卷壁方降低了c-jun和p65的表达,表明卷壁方抑制了RA中AP-1和NF-κB通路的表达。
■卷壁方的核心活性分子可以抑制AP-1和NF-κB通路的关键因子,从而抑制炎症,对RA有保护作用。