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目的:观察血小板对大鼠类风湿关节炎滑膜炎症反应和细胞增生的影响,探讨蛭元方抑制血小板加剧类风湿关节炎滑膜细胞炎症和增生反应的可能的分子机制。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、血小板对照组、类风湿关节炎滑膜模型组、血小板干预组和蛭元方干预组。在不同浓度TNF诱发的大鼠类风湿关节炎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类风湿关节炎模型细胞,采用ELISA法分别检测RSC364细胞培养上清中IL-6、IL-8的生成量和P-选择素的表达情况,采用MTT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TNF存在下,类风湿关节炎滑膜模型组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与类风湿关节炎滑膜模型组相比,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:活化血小板能加剧类风湿关节炎滑膜细胞的炎症和增生反应;蛭元方能有效缓解血小板对大鼠类风湿关节炎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。
OBJECTIVE: To observe the effect of platelet on the synovial inflammatory reaction and cell proliferation in rats with rheumatoid arthritis, and to explore the possible molecular mechanism of leiyuanfang inhibiting platelets to aggravate the inflammation and proliferation of rheumatoid arthritis synovial cells. Methods: The routine cultured rat fibroblast-like synovial cell line RSC364 was randomly divided into normal control group, platelet control group, rheumatoid arthritis synovial model group, platelet intervention group and lee-Yuan Fang intervention group. In different concentrations of TNF-induced rat rheumatoid arthritis synovial cell model based on bovine type II collagen stimulated platelet activation with serum containing Hirudoe prescription activated platelet intervention in rheumatoid arthritis model cells The production of IL-6 and IL-8 and the expression of P-selectin in the culture supernatant of RSC364 cells were detected by ELISA. The proliferation of RSC364 cells was observed by MTT assay. Results: (1) When platelet of bovine type II collagen activated platelets 5h, platelets expressed the most P-selectin; (2) IL-6 and IL-8 in the supernatant of RSC364 cells of rheumatoid arthritis synovial model group (P <0.05). The proliferation rate of RSC364 cells was also significantly higher than that of the normal control group (P <0.05), and showed a dose-dependent manner; ③ Compared with rheumatoid arthritis synovial model group, the amount of IL-6 and IL-8 produced by RSC364 cells in platelet intervention group increased significantly (P <0.05 and P <0.01) and the proliferation rate of synovial cells increased significantly (P <0.05) .④Compared with platelet intervention group, the production of IL-6 and IL-8 of RSC364 cells in the treatment group was significantly decreased (P <0.05 and P <0.01), and the proliferation rate was also significantly decreased (P <0.05), while P-selectin expression was significantly reduced (P <0.05). Conclusion: Activated platelets can aggravate the inflammation and proliferation of rheumatoid arthritis synovial cells; leech can effectively alleviate the role of platelets on the synovial inflammation and proliferation of rheumatoid arthritis in rats, the mechanism may be related to inhibition of P-select The expression of prime.