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目的:研究京尼平苷酸(GA)对佐剂性关节炎(AA)模型大鼠滑膜细胞体外培养增殖能力和分泌细胞因子的影响。方法:用弗氏完全佐剂建立AA模型大鼠,分离其滑膜细胞进行培养,以阳性细胞数和纯度鉴定其免疫细胞;将细胞分为对照组,GA低、中、高(10-7、10-6、10-5mol·L-1)剂量组,阳性对照(甲氨蝶呤,10-6mol·L-1)组并加入相应药物处理。MTT法检测吸光度(A)考察GA对滑膜细胞增殖的影响;流式细胞术测定细胞周期;酶联免疫吸附法检测各组滑膜细胞分泌肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-10的水平。结果:2～3代滑膜细胞纯度达到试验要求;与对照组比较,GA高剂量组和阳性对照组大鼠滑膜细胞A值,S和G2/M期细胞比例,细胞外液TNF-α、IL-1β水平明显降低(P<0.05或P<0.01),G1期细胞比例、IL-10水平明显升高(P<0.05或P<0.01)。结论:GA能显著抑制AA模型大鼠滑膜细胞的增殖,阻滞细胞周期于G1期,抑制滑膜细胞分泌TNF-α和IL-1β,促进IL-10分泌。
AIM: To investigate the effects of geniposidic acid (GA) on the proliferation and cytokine secretion of synoviocytes cultured in vitro in rats with adjuvant arthritis (AA). Methods: AA model rats were established by complete Freund ’s adjuvant. The synovial cells were isolated and cultured. The number of immunocytes was identified by the number of positive cells and purity. The cells were divided into control group, GA low, middle and high (10-7 , 10-6,10-5mol·L-1) dose group, positive control group (methotrexate, 10-6mol·L-1) group and add the appropriate drug treatment. MTT assay absorbance (A) to investigate the effect of GA on the proliferation of synovial cells; flow cytometry was used to determine the cell cycle; enzyme-linked immunosorbent assay was used to detect the levels of tumor necrosis factor-α (TNF-α), interleukin -1 (IL-1β), IL-10. Results: The purity of synovial cells of 2 ~ 3 generations reached the requirement of experiment. Compared with the control group, the content of synoviocyte A, the proportion of cells in S and G2 / M phase, the level of TNF-α in extracellular fluid (P <0.05 or P <0.01). The percentage of cells in the G1 phase and the level of IL-10 were significantly increased (P <0.05 or P <0.01). CONCLUSION: GA can significantly inhibit the proliferation of synovial cells in AA model rats, arrest the cell cycle in G1 phase, inhibit the secretion of TNF-α and IL-1β by synovial cells and promote the secretion of IL-10.