Abstract
Purpose : To investigate the effect and mechanism of action of Ermiao san (EMS), a traditional Chinese herbal formula, on inflammation development and production of inflammatory mediators in adjuvantinduced arthritis (AIA). Methods : AIA was induced by injection of 0.1 ml Freund’s complete adjuvant (FCA, 10 mg/ml) in the left hind footpad of the rats. AIA rats were intragastricly treated with 0.5, 1, 2 g/kg EMS or 0.1 g/kg methotrexate from day 7 to 28 after FCA challenge. Foot volume and histological score were measured. Osteoclast number was calculated by tartrate-resistant acid phosphatase (TRAP) staining assay. Levels of prostaglandin (PG) E2, tumor necrosis factor (TNF) -α and interleukin (IL)-1β in serum were determined by enzyme-linked immunosorbent assay (ELISA) while the level of nitric oxide (NO) in serum was analyzed by Griess reaction method. Results : Foot volume, histological score, osteoclast number and serum levels of TNF-α, IL-1β, PGE2 and NO were all increased in AIA group rats on day 28 after FCA challenge (all p < 0.01) compared with control. EMS (1 and 2 g/kg) significantly decreased the foot volume of AIA rats by 10 % (p < 0.05) and 19 % (p < 0.01), respectively, compared with AIA group. Furthermore, 1 and 2 g/kg EMS significantly reduced histological score by about 28 % (p < 0.05) and 46 % (p < 0.01), respectively, as well as osteoclast number by 12 % (p < 0.05) and 15 % (p < 0.05), respectively, compared with AIA group. In addition, 1 and 2 g/kg EMS significantly decreased the serum levels of TNF-α about 23 % (p < 0.05) and 43 % (p < 0.01), IL-1β by15 % (p < 0.05) and 26 % (p < 0.01), NO 13 % (p < 0.05) and 26 % (p < 0.01) as well as PGE2 by 11 % (p < 0.05) and 15 % (p < 0.01), respectively, compared with AIA group. Conclusion : These results suggest that EMS probably alleviates arthritis development and joint destruction by decreasing the production of inflammatory mediators in AIA rats. Keywords : Ermiao san, Inflammatory mediators, Adjuvant-induced arthritis, Traditional Chinese medicine, Rheumatoid arthritis
Highlights
Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation with the lesions of cartilage and bone, and results in joint destruction and deformity [1]
On treatment with 1 and 2 g/kg Ermiao san (EMS) for 21 days, foot volume of arthritic rats were significantly decreased compared with adjuvant-induced arthritis (AIA) group given saline (p < 0.05 and p < 0.01 respectively, Figure 1)
Treatment with EMS significantly lowered the levels of tumor necrosis factor (TNF)-α, IL-1β, prostaglandin E2 (PGE2) and nitric oxide (NO) in serum of AIA rats, the degree of reduction were lower than methotrexate, the positive controlled drug used in this study
Summary
Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation with the lesions of cartilage and bone, and results in joint destruction and deformity [1]. In the present study, we investigated the effect of EMS on the arthritis development and production of inflammatory mediators in AIA of rats. Herbarium voucher specimens of the tested herbs were deposited at the Chongqing Key Laboratory of Nature Medicine Research, Chongqing Technology and Business University, with voucher specimen numbers as follows: 2013-109 (Atractylodes chinensis (DC.) Koidz.) and 2013-187 268 g EMS freeze-dried extract powder was obtained from 1000 g raw material (the yield was 26.8 %). Levels of TNF-α, IL-1β and PGE2 in serum were tested using enzyme-linked immunosorbent kits. Histopathological changes in joint were scored (histological score) using the following parameters by one investigator blind to the experimental protocol [15]: 0: normal, 1: infiltration of inflammatory cells, 2: synovial hyperplasia, 3: pannus formation, 4: bone erosion, 5: bone destruction. Differences between values were considered significant at p < 0.05
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