Methotrexate And Alkylating Agents For Rheumatoid Arthritis Peripheral Blood Lymphocytes To Growth, Cell Cycle And The Impact Of Ifn-¦Ã And Il-4 Secretion Levels

Object To explore the effect of methotrexate(MTX) and alkylatingagent(chlorethazine is selected) on cell proliferation, cell cycle and cytokines ofthe peripheral blood lymphocyte in patients with rheumatoid arthritis, toinvestigate its possible synergistic effect and evaluate the possible mechanisms. Method The level of cell proliferation was measured by the MTTmethod, cell cycle was measured by flow cytometry with PI Staining (FCM)method, and the level of IFN-γ and IL-4 was measured with ELISA. Result Level of proliferation of MTX-treated (2.5 and 5mg/L) andHN2-treated (0.025 and 0.25mg/L) PBL in patients with RA decreasedsignificantly compared with control group, level of proliferation ofMTX+HN2-treated PBL decreased compared with the correspondingsingle-drug-treated groups significantly(p<0.05);FCM assay showed the rateof MTX-treated PBL in G₀/G₁ phase increased but the rate of MTX-treated PBLin S phase decreased compared with control group; the rate of HN₂-treated PBLin G₀/G₁ phase increased but the rate of HN₂-treated PBL in S and G₂ /M phasedecreased compared with control group; the rate of MTX +HN2-treated PBL inG₀/G₁ phase increased and the rate of MTX+HN₂-treated PBL in S and G₂/Mphase decreased more obviously. Furthermore, 2.5 mg/L MTX and 0.25mg/LHN₂ augmented apoptosis of PBL. level of IFN-γ of MTX-treated andHN₂-treated groups dropped significantly compared with control group, and thatof MTX+HN2-treated group dropped significantly compared with thecorresponding single-drug-treated groups(p<0.05); but all of the groups had noeffect in the production of IL-4 in these doses(p>0.05). Conclusion The findings suggest the combined therapy(MTX+HN₂) hasthe synergistic effect in inhibiting the proliferation, proliferation-dependedprocesses and the production of IFN-γ of PBL with RA patients.