摘要
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1.0μmol/L组、白花丹醌1.5μmol/L组、白花丹醌2.0μmol/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-α、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。
Objective To observe the effects of plumbagin on proliferation and the expressions of tumor necrosis factor-α （ TNF-α）, interleukin-1 β （ IL-1 β） 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-α, 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 MTF 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-1 βand 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.
作者
钟毓娟
李丽
李勇文
杨成芳
方舒萍
程琪
Zhong Yujuan1 , Li Li2, Li Yongwen3, et al(1Dept oflmmunology,2Dept of Physiology,3Dept of Pharmacology, Guilin Medical University, Guilin 541001)
出处
《安徽医科大学学报》
CAS
北大核心
2018年第3期343-347,共5页
Acta Universitatis Medicinalis Anhui
基金
广西高校科学技术研究项目(编号:KY2015LX279)
