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目的观察清热活血方治疗活动期类风湿关节炎的可能作用机制。方法分离类风湿关节炎患者的滑膜组织制备原代类风湿关节炎成纤维样滑膜细胞(RA-FLS),采用肿瘤坏死因子α(TNF-α)诱导RA-FLS的迁移和黏附。设空白组(不加药物及TNF-α干预)、诱导组(10或20 ng/ml TNF-α诱导)、清热活血方低、中、高剂量组(20、100、500 ng/ml清热活血方+TNF-α诱导)。检测各组细胞迁移距离及面积、细胞黏附OD值和黏附抑制率,并检测细胞中PI3K/Akt信号通路相关蛋白(PI3K、p-Akt、Akt)表达。结果与空白组比较,诱导组细胞迁移距离及面积增加、黏附OD值增大、PI3K及p-Akt蛋白表达升高(P<0.01);与诱导组比较,清热活血方各剂量组均可抑制细胞的迁移和黏附以及PI3K、p-Akt蛋白表达,并以500 ng/ml效果最好(P<0.05或P<0.01)。结论清热活血方可能通过下调RA-FLS中PI3K、p-Akt蛋白表达,抑制RA-FLS的迁移和黏附,从而减缓滑膜炎症和关节破坏的进展。
Objective To observe the possible mechanism of Qingre Huoxue Recipe in the treatment of active rheumatoid arthritis. Methods Primary rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) were isolated from synovial tissue of patients with rheumatoid arthritis. Tumor necrosis factor-α (TNF-α) was used to induce RA-FLS migration and adhesion. The rats in the blank group (without drug and TNF-α intervention), the induction group (induced by 10 or 20 ng / ml TNF-α), the rats in the low, medium and high dose of heat- Fang + TNF-α induction). The cell migration distance and area, cell adhesion OD value and adhesion inhibition rate were detected. The expression of PI3K / Akt signal pathway related proteins (PI3K, p-Akt, Akt) were detected. Results Compared with the blank group, the migrating distance and area of the induced group increased, the OD value of the adhesion increased, and the expression of PI3K and p-Akt protein increased (P <0.01). Compared with the induction group, The migration and adhesion of cells and the expression of PI3K and p-Akt protein showed the best effect at 500 ng / ml (P <0.05 or P <0.01). Conclusion Qingre Huoxue Prescription can reduce the synovial inflammation and the progression of joint destruction by decreasing the expression of PI3K and p-Akt in RA-FLS and inhibiting the migration and adhesion of RA-FLS.