Shu Haiyang, Zhao Hanxiao, Shi Yingjie, Lu Cheng, Li Li, Zhao Ning, Lu Aiping, He Xiaojuan
The Second Clinical College, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China.
Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
Chin Med. 2021 Apr 12;16(1):31. doi: 10.1186/s13020-021-00439-w.
Rheumatoid arthritis (RA) is a chronic autoimmune disease accompanied with joint destruction that often leads to disability. Wang-Bi capsule (WB), a traditional Chinese medicine-based herbs formula, has exhibited inhibition effect on joint destruction of collagen-induced arthritis (CIA) animal model in our previous study. But its molecular mechanisms are still obscure.
CIA rats were treated intragastrical with WB for eight weeks, and the effect of joints protection were evaluated by hematoxylin and eosin (H&E) staining, safranin O fast green staining, tartrate-resistant acid phosphatase (TRAP) staining and micro‑CT scanning analysis. The transcriptomic of tarsal joints were used to investigate how WB alleviated joint destruction.
The histological examination of ankle joints showed WB alleviated both cartilage damage and bone destruction of CIA rats. This protective effect on joints were further evidenced by micro-CT analysis. The transcriptomic analysis showed that WB prominently changed 12 KEGG signaling pathways ("calcium signaling pathway", "cAMP signaling pathway", "cell adhesion molecules", "chemokine signaling pathway", "complement and coagulation cascades", "MAPK signaling pathway", "NF-kappa B signaling pathway", "osteoclast differentiation", "PI3K-Akt signaling pathway", "focal adhesion", "Gap junction" and "Rap1 signaling pathway") associated with bone or cartilage. Several genes (including Il6, Tnfsf11, Ffar2, Plg, Tnfrsf11b, Fgf4, Fpr1, Siglec1, Vegfd, Cldn1, Cxcl13, Chad, Arrb2, Fgf9, Egfr) regulating bone resorption, bone formation and cartilage development were identified by further analysis. Meanwhile, these differentially expressed genes were validated by real-time quantitative PCR.
Overall, the protective effect of WB treatment on joint were confirmed in CIA rats, and its basic molecular mechanisms may be associated with regulating some genes (including Il6, Tnfsf11, Ffar2 and Plg etc.) involved in bone resorption, bone formation and cartilage development.
类风湿关节炎（RA）是一种慢性自身免疫性疾病，常伴有关节破坏，进而导致残疾。在我们之前的研究中，基于中药的方剂——尪痹胶囊（WB）对胶原诱导性关节炎（CIA）动物模型的关节破坏具有抑制作用。但其分子机制仍不清楚。
用WB灌胃治疗CIA大鼠8周，通过苏木精-伊红（H&E）染色、番红O固绿染色、抗酒石酸酸性磷酸酶（TRAP）染色和显微CT扫描分析评估关节保护效果。利用跗关节转录组研究WB如何减轻关节破坏。
踝关节组织学检查显示，WB减轻了CIA大鼠的软骨损伤和骨质破坏。显微CT分析进一步证明了其对关节的这种保护作用。转录组分析表明，WB显著改变了12条与骨骼或软骨相关的KEGG信号通路（“钙信号通路”“cAMP信号通路”“细胞黏附分子”“趋化因子信号通路”“补体和凝血级联反应”“丝裂原活化蛋白激酶信号通路”“核因子κB信号通路”“破骨细胞分化”“磷脂酰肌醇3激酶-蛋白激酶B信号通路”“黏着斑”“缝隙连接”和“Rap1信号通路”）。通过进一步分析，鉴定出了几个调节骨吸收、骨形成和软骨发育的基因（包括Il6、Tnfsf11、Ffar2、Plg、Tnfrsf11b、Fgf4、Fpr1、Siglec1、Vegfd、Cldn1、Cxcl13、Chad、Arrb2、Fgf9、Egfr）。同时，通过实时定量PCR验证了这些差异表达基因。
总体而言，在CIA大鼠中证实了WB治疗对关节的保护作用，其基本分子机制可能与调节一些参与骨吸收、骨形成和软骨发育的基因（包括Il6、Tnfsf11、Ffar2和Plg等）有关。
