The Application Of "Qu Feng Qing Re"-Expelling Wind And Clearing Away Heat And "Huo Xue Tong Luo"-Promoting Blood Circulation To Remove Meridian Ob-Struction Method In Rheumatoid Arthritis

Objective: To establish a formula Jin Bian Qu Feng Yin (JBY) in treatment ofrheumatoid arthritis based on the theory of traditional Chinense medicine “Qu FengQing Re”-expelling wind and clearing away heat and “Huo Xue Tong Luo”-promoting blood circulation to remove meridian obstruction. To observe the influenceof JBY liquid on production of proinflammatory cytokines and metalloproteinases incultured synovial fibroblasts from patients with rheumatoid arthritis, evaluatingpossible mechanisms of JBY in treatment of rheumatoid arthritis.Methods: The synovial fluid is collected from6patients with rheumatoid arthritis(RA), as well as from6patients with knee osteoarthritis (OA) as control. The cells insynovial fluid are cultured in25cm2bottles and stimulated with low, middle, highdose of JBY, and the level of proinflammatory cytokines IL-1β, TNF-α andmetalloproteinase MMP-3are detected by ELISA and RT-PCR. The proliferation ofthe cells is evaluated by MTT.Results:(1) Synovial cell morphological change: after repeated attached wall andpassaged, synovial cells gradually purified as fibroblast-like cells. After stimulated byJBY72hours, the cells treated with middle or high dose of JBY became small.However, the cells treated with low dose JBY slightly changed as multi-form. Theproliferation of synovial fibroblasts was significantly interfered by JBY with the dosedependent manner；(2) Measured by ELISA the culture supernatant of InflammatoryCytokines: Stimulated72h with different concentration of JBY, culture supernatantsof TNF-ɑ, IL-1β MMP-3were lower, with the OA control group compared significantdifferences (P <0.05), has a very significant difference (P <0.01) compared with thecontrol group,between the two groups, the high dose group in reducing TNF-α, IL-1βand MMP-3level, a significant difference (P <0.05)；(3) MTT assay of RA and OAsynovial cell proliferation:Stimulated72h with different concentration of JBY, thelevel of TNF-α, IL-1β MMP-3in supernatant of synovial cells reduced significantlywith the dose dependent manner, lower in middle-and high-dose JBY-stimulated cellsthan in low-dose stimulated cells.(P <0.05)；(4) RT-PCR show RA synovial cells ofTNF-α, IL-1β and MMP-3expression:In the cells the mRNA expression level ofTNF-α, IL-1β and MMP-3also reduced after co-cultured by JBY. These results weresuggested that JBY could inhibit production of proinflammatory cytokines and MMPsin synovial cells.Conclusion: Based on the theory of traditional Chinese medicine “expelling wind and removing heat and promoting blood circulation to remove meridian obstruction”, theformula JBY significantly suppress RA synovial cell proliferation and reduce IL-1β,TNF-α and MMPs-3production, suggesting JBY may play important role in treatmentof rheumatoid arthritis. The mechanism of the traditional Chinense medical formulashould be further studied.