Inflammation and Immunology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto 602-8566, Japan.
J Clin Biochem Nutr. 2008 Sep;43(2):126-8. doi: 10.3164/jcbn.2008057.
Eicosapentaenoic acid (EPA) is essential for normal cell growth, and may play an important role in inflammatory and autoimmune disorders including rheumatoid arthritis. We investigate that EPA could suppress the proliferation of fibroblast like synoviocytes in vitro. We treated synoviocytes with 1 to 50 microM EPA and measured cell viabilities by the modified MTT assay. We sorted the number of them in sub G1 stage by fluorescence-activated cell sorting caliber. And we stained them by light green or Hoechst 33258, and investigate microscopic appearance. The cell viabilities were decreased at 30 microM, 40 microM, and 50 microM of EPA comparing to 0 microM of EPA. The half maximal concentration of synoviocytes inhibition was approximately 25 microM. At day 1 and day 3, cell number was also decreased at 50 microM EPA comparing to control. FACS caliber indicated the number of synoviocytes in sub G1 stage did not increase in each concentration of EPA. Hoechst staining indicated normal chromatin pattern and no change in a nuclear morphology both in EPA treated synoviocytes and in untreated synoviocytes. These findings suggest that EPA could suppress the proliferation of synoviocytes in vivo dose dependently and time dependently, however, the mechanism is not due to apoptosis.
二十碳五烯酸(EPA)是正常细胞生长所必需的,并且可能在包括类风湿性关节炎在内的炎症和自身免疫性疾病中发挥重要作用。我们研究了 EPA 可以抑制体外成纤维样滑膜细胞的增殖。我们用 1 到 50 μM 的 EPA 处理滑膜细胞,并通过改良的 MTT 测定法测量细胞活力。我们通过荧光激活细胞分选仪(FACS)测量亚 G1 期细胞的数量。我们用亮绿或 Hoechst 33258 染色,观察微观形态。与 0 μM EPA 相比,在 30 μM、40 μM 和 50 μM EPA 下细胞活力降低。半数最大抑制浓度的滑膜细胞约为 25 μM。在第 1 天和第 3 天,与对照相比,50 μM EPA 组的细胞数量也减少了。FACS 分析表明,在 EPA 的每个浓度下,亚 G1 期的滑膜细胞数量并没有增加。Hoechst 染色表明,在 EPA 处理的滑膜细胞和未处理的滑膜细胞中,染色质形态正常,核形态没有变化。这些发现表明,EPA 可以在体内剂量依赖性和时间依赖性地抑制滑膜细胞的增殖,但机制不是由于细胞凋亡。