蛭元方对血小板加剧的类风湿关节炎滑膜炎症和增生的影响

目的:观察蛭元方对血小板加剧的类风湿关节炎( rheumatoid arthritis,RA)大鼠滑膜细胞炎症和增生反应的影响.方法:将常规培养的大鼠类纤维母样滑膜细胞株RSC364随机分为正常对照组、RA模型组、活化血小板干预组和蛭元方干预组.在不同浓度TNF-α诱发的大鼠RA滑膜细胞模型的基础上,以牛Ⅱ型胶原蛋白刺激血小板活化,用蛭元方含药血清作用于活化血小板干预下的RA模型细胞,采用酶联免疫吸附试验(ELISA)法分别检测RSC364细胞培养上清中白细胞介素-6(IL-6)、白细胞介素-8(IL-8)的生成量和P-选择素的表达情况,采用MTT比色法观察RSC364细胞增殖的情况.结果:①牛Ⅱ型胶原蛋白活化血小板5h时,血小板表达P-选择素最多;②在不同浓度肿瘤坏死因子-α(TNF-α)存在下,RA模型组RSC364细胞培养上清中IL-6和IL-8的生成量均增加,与同一浓度的正常对照组相比均有显著性差异(P＜0.05);RSC364细胞增殖率也明显高于正常对照组(P＜0.05),并呈现剂量依赖性;③与RA模型组相比,活化血小板干预组RSC364细胞生成IL-6和IL-8的量均明显增加(P＜0.05和P＜0.01),滑膜细胞增殖率显著升高(P＜0.05);④与血小板干预组相比,蛭元方干预组RSC364细胞的IL-6和IL-8生成量均显著降低(P＜0.05和P＜0.01),增殖率亦有明显下降(P＜0.05),同时P-选择素的表达量显著降低(P＜0.05).结论:活化血小板能加剧RA滑膜细胞的炎症和增生反应;蛭元方能有效缓解血小板对RA大鼠滑膜炎症及增生的促进作用,其机制可能与抑制P-选择素的表达有关.

Effects of Zhiyuanfang on Activated Platelets Amplify Inflammation and Proliferation of Synoviocytes in Rheumatoid Arthritis

Objective:To observe the effects of Zhiyuanfang inhibiting platelets amplified inflammation and proliferation of synovial cells in the rheumatoid arthritis(RA) rats.Method:Normal cultured RSC364 cells were divided into normal control group,RA model group,activated platelet intervention group and Zhiyuanfang group.The different concentrations of tumor necrosis factor-α(TNF-α) were used to induce RA rat cell models,stimulate platelets activating by bovine type II collagen,and interfere platelet intervention group with Zhiyuanfang containing serum.The expression of interleukin-6(IL-6),interleukin-8(IL-8) and P-selecting were assayed respectively by enzyme-linked immunosorbnent assay(ELISA),and the proliferation of RSC364 cells were determined by MTT colorimetric assay.Result:①The expression of P-selecting was at peak when platelets were activated by bovine type II collagen for 5 h.②the expression of IL-6 and IL-8 on synovial cells in RA model group was increased,which was compared with the same concentration of normal group showed a significant difference(P<0.05);the rate of RSC364 cells proliferation was also higher than normal group(P<0.05)in a dose-dependent manner.③Compared with RA model group,the expression of IL-6 and IL-8 on synovial cells in platelets intervention group was significantly increased(P<0.05,P<0.01),and the rate of synovial cells proliferation was also higher(P<0.05).④ Compared with platelets intervention group,the expression of IL-6 and IL-8 on synovial cells in Zhiyuanfang group was significantly reduced(P<0.05,P<0.01),the proliferation rate of synovial cells was declined(P<0.05),and the expression of P-selecting was also lower.Conclusion:Activated platelets amplify the inflammation and proliferation of synoviocytes;Zhiyuanfang effectively can relieve the amplification of platelets on the inflammation and proliferation in RA rats probably by the inhibition of P-selecting expression.