苦参碱对类风湿关节炎外周血淋巴细胞生长、细胞周期及IFN-γ、IL-4水平影响的研究

【作者】 王晓丽；

【导师】 李小峰；

【作者基本信息】 山西医科大学 ， 风湿免疫学， 2007， 硕士

【摘要】 目的在体外实验中,通过比较中药苦参碱（Mat Matrine）与甲氨蝶呤（MTX Methotrexate）对类风湿关节炎患者外周血淋巴细胞（PBL）生长、细胞周期及IFN-γ与IL-4分泌水平的影响,探讨Mat对类风湿关节炎PBL的作用机制。方法1.分离纯化外周血淋巴细胞,进行体外培养,选取MTX和Mat两种药物,各设6个浓度,其中O浓度为对照组。2.应用MTT比色法测定24小时和48小时的MTX处理组、Mat处理组的吸光度值（A）,计算细胞抑制率。3.应用碘化丙啶（PI）染色流式细胞术（FCM）检测48小时MTX处理组、Mat处理组的细胞周期;4.应用酶联免疫吸附试验（ELISA法）检测MTX处理组、Mat处理组的IFN-γ与IL-4的分泌水平。结果1.MTX、Mat均可抑制体外培养的PBL生长,随着药物浓度的增大,吸光度值（A）下降,细胞抑制率（IR%）增加,呈剂量依赖性。48h作用点细胞抑制率（IR%）明显大于24h作用点（P＜0.001）。两药物比较无明显差别（P＞0.05）。2.MTX、Mat均可使细胞周期构成发生明显变化,与对照组相比存在显著差异（P＜0.001）,G₀/G₁期细胞比例增高,S期和G₂/M期细胞比例降低。随着药物浓度增加,细胞增殖指数逐渐降低,呈剂量依赖性。两药物比较无明显差别（P＞0.05）。3.MTX、Mat均可降低促炎因子IFN-γ分泌水平,与对照组相比存在显著差别（P＜0.001）,但Mat组IFN-γ分泌水平降低幅度小于MTX组（P＜0.001）。MTX、Mat组IL-4分泌水平与对照组无明显差别（P＞0.05）。结论1.Mat抑制体外培养的RA患者PBL生长的作用与MTX相当。2.Mat对PBL细胞周期的阻滞作用与MTX相当。3.与MTX相比,Mat可下调IFN-γ分泌水平,Mat作用弱于MTX。MTX、Mat对IL-4分泌水平无明显影响。4.Mat、MTX作用机制有相似性,推测Mat可能具有治疗RA的潜在价值。更多还原

【Abstract】 ObjectiveTo investigate the effects of Matrine（Mat）on cell proliferation,cell cycling and cytokines of the peripheral blood lymphocyte in patients with rheumatoid arthritis.Furthermore,through contrast with methotrexate（MTX）,try to find out the mechanism of treatment for rheumatoid arthritis（RA）.Method1.Separate and purify lymphocyte cells.Culture lymphocyte cells outside of body.Choose Methotrexate and Matrine.2.The level of cell proliferation was measured by the MTT method.3.Cell cycle was measured by flow cytometry with PI Staining（FCM）method.4.The level of IFN-γand IL-4 was measured with ELISA.Result1.Both MTX and Mat can inhibit the proliferation of PBL in patients with RA and the effect is dose-dependent.It makes IR%increasing.（P＜0.05）.IR%of 48h is more than IR%of 24h（P＜0.001）.The two medicine have no obvious difference（P＞0.05）.2.Both MTX and Mat can change the cell cycle and act in G₁/S point.They both make the number of cells in G₀/G₁ phase increasing and cells in S and G₂/M phase decreasing（P＜0.05）. The two medicine have no obvious difference（P＞0.05）.3.Level of IFN-γof Mat-treated and MTX-treated groups dropped significantly compared with control group（P＜0.001）.But the level of IFN-γof Mat-treated groups is more than MTX -treated groups.All of the groups had no effect in the production of IL-4 in these doses （P＞0.05）.Conclusion1.Mat inhibites the proliferation of PBL in patients with RA.The effect is correspondent to MTX.2.Mat inhibites proliferation-dependent processes of PBL in patients with RA.The effect is correspondent to MTX.3.The effect on IFN-γof Mat is weaker than MTX.4.The mechanism of Mat is similar to MTX.Mat maybe have the potential value in the treatment of RA.更多还原