School of Pharmacy, Anhui University of Chinese Medicine, Hefei, China.
Anhui Province Key Laboratory of Chinese Medicinal Formula, Hefei, China.
Pharm Biol. 2022 Dec;60(1):846-853. doi: 10.1080/13880209.2022.2066700.
Er Miao San (EMS) is a formulation that contains and in 1:1 ratio, and is commonly used to treat rheumatoid arthritis (RA) and other inflammatory diseases.
We investigated the mechanism of action and effects of EMS on peritoneal macrophage differentiation in a rat model of adjuvant arthritis (AA).
EMS (3, 1.5 and 0.75 g/kg; once daily) and methotrexate (0.5 mg/kg; once every 3 days) were administered orally from days 21 to 35 after immunisation. Paw swelling and arthritis index were measured; pathological changes in the ankle joint were observed using x-ray and haematoxylin eosin staining. The ratio of CD86/CD206 in macrophages was detected by flow cytometry. Examination of the miRNA-33/NLRP3 signalling pathway was examined by RT-qPCR and western blotting. The levels of cytokines in the serum and cell supernatants were tested by ELISA.
EMS significantly reduced the AA index in rats (from 11.0 to 9.3) and pathological changes in the ankle joint (from 3.8 to 1.4). The ratio of CD86/CD206 was reduced, and polarisation to M1 improved (from 0.9 to 0.6) in macrophages of EMS-treated rats. EMS downregulated the miRNA-33/NLRP3 pathway. Furthermore, EMS treatment increased IL-10 and TGF-β levels in the serum and supernatant of macrophages of AA rats and simultaneously decreased the levels of IL-1β and TNF-α.
Our results suggest that EMS may reduce macrophage polarisation to the M1 inflammatory phenotype by downregulating the miRNA-33/NLRP3 pathway in AA rats. These findings may provide new insights into the treatment of RA.
二妙散（EMS）是一种含有 和 的配方，以 1:1 的比例混合，常用于治疗类风湿关节炎（RA）和其他炎症性疾病。
我们研究了 EMS 在佐剂性关节炎（AA）大鼠模型中对腹腔巨噬细胞分化的作用机制和效果。
从免疫后第 21 天到第 35 天，每天口服 EMS（3、1.5 和 0.75 g/kg；一次）和甲氨蝶呤（0.5 mg/kg；每 3 天一次）。测量爪肿胀和关节炎指数；通过 X 射线和苏木精-伊红染色观察踝关节的病理变化。通过流式细胞术检测巨噬细胞中 CD86/CD206 的比值。通过 RT-qPCR 和 Western blot 检测 miRNA-33/NLRP3 信号通路。通过 ELISA 检测血清和细胞上清液中细胞因子的水平。
EMS 显著降低了 AA 大鼠的 AA 指数（从 11.0 降至 9.3）和踝关节的病理变化（从 3.8 降至 1.4）。EMS 处理的大鼠巨噬细胞中 CD86/CD206 的比值降低，向 M1 极化改善（从 0.9 降至 0.6）。EMS 下调了 miRNA-33/NLRP3 通路。此外，EMS 治疗增加了 AA 大鼠血清和巨噬细胞上清液中 IL-10 和 TGF-β 的水平，同时降低了 IL-1β 和 TNF-α的水平。
我们的结果表明，EMS 可能通过下调 AA 大鼠的 miRNA-33/NLRP3 通路，减少巨噬细胞向 M1 炎症表型的极化。这些发现可能为 RA 的治疗提供新的思路。
