Objective To observe the effects of plumbagin on proliferation and the expressions of tumor necrosis factor-a(TNF-α), interleukin-1β (IL-1β) and matrix metalloproteinase-3 (MMP-3) in fibroblast-like synoviocytes(FLS) induced by lipopolysaccharide(LPS). Methods Using type II collagen and Freund's complete adjuvant to make arthritis model in rat. CIA-FLS were primary cultured in vitro, the effect of drug on proliferation of CIA-FLS was detected by MTT method. CIA-FLS were divided into CIA control group, LPS control group, methotrexate group, three concentration groups of plumbagin 1.0, 1.5 or 2. 0 μmol/L; after being incubated with each drug for 48 hours, the levels of TNF-a, IL-1β and MMP-3 in cell culture supernatant were assayed by ELISA; the protein content of MMP-3 in CIA-FLS was mesured by Western blot. Results The result of MTT showed that plumbagin could inhibit the proliferation of CIA-FLS in different degrees, presenting in a concentration and time dependence manner. Compared with LPS control group, the contents of TNF-α, IL-1β and MMP-3 in cell culture supernatant were significantly decreased in methotrexate, the 1.0, 1.5 and 2. 0 μmol/L of plumbagin groups (P＜ 0. 001); methotrexate or plumbagin could also significantly reduce the protein expression of MMP-3 in CIA-FLS (P＜ 0. 001). Conclusion Plumbagin can inhibit the proliferation of CIA-FLS and the inflammatory reaction induced by LPS via decreasing the expressions of TNF-α, IL-1β and MMP-3.%目的 研究白花丹醌对脂多糖(LPS)诱导的体外培养CIA大鼠成纤维样滑膜细胞(CIA-FLS)的增殖及肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)和基质金属蛋白酶-3(MMP-3)表达的影响.方法 采用Ⅱ型胶原蛋白+弗氏完全佐剂建立大鼠关节炎模型(CIA);原代培养CIA-FLS,MTT法检测药物对CIA-FLS增殖的影响;随机设立CIA对照组、 LPS对照组、甲氨蝶呤组、白花丹醌1.0 μmol/L组、白花丹醌1.5 μmol/L组、白花丹醌2.0 μmol/L组;各药物与细胞共同孵育48 h后,ELISA法检测细胞培养上清液中TNF-a、 IL-1β和MMP-3的含量;Western blot检测细胞内MMP-3的蛋白含量.结果 MTT显示,白花丹醌能不同程度抑制 CIA-FLS的增殖,呈浓度和时间依赖性;与LPS对照组比较,甲氨蝶呤或白花丹醌均能明显下调细胞培养上清液中TNF-α、 IL-1β和MMP-3的含量(P<0. 001),明显降低细胞中 MMP-3的蛋白含量(P<0.001).结论 白花丹醌能抑制 CIA-FLS的增殖,并能通过下调TNF-α、IL-1β和MMP-3的表达而抑制LPS诱导CIA-FLS的炎症反应.
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