紫草素对类风湿关节炎滑膜成纤维细胞COX-2表达的影响

Effect of shikonin on the COX-2 expression of synovial fibroblasts in rheumatoid arthritis

目的观察紫草素对人滑膜成纤维细胞环氧化酶-2(COX-2)mRNA表达的影响,探讨紫草素可能的免疫抗炎作用机制。方法分离并培养人膝关节滑膜成纤维细胞,各加入IL-1β、TNF-α15 ng/mL,体外模拟人类风湿关节炎滑膜细胞。以塞来昔布作阳性对照,进行紫草素(浓度为5～80μg/L)干预,分别作用24、48、72 h后终止培养。用半定量逆转录-聚合酶链反应(RT-PCR)技术检测滑膜成纤维细胞COX-2 mRNA表达情况。结果紫草素对滑膜成纤维细胞COX-2 mRNA的表达,对浓度无明显依赖性(P=0.089),用药时间上有显著统计学意义(P<0.0001)。塞来昔布抑制滑膜成纤维细胞COX-2mRNA水平的表达亦无明显浓度依赖性(P=0.051),而与用药时间显著相关(P<0.0001)。紫草素对滑膜成纤维细胞COX-2 mRNA表达的影响与塞来昔布相比,其抑制作用无明显差异。结论紫草素可通过抑制COX-2 mRNA基因表达,减少炎症性细胞因子的产生而发挥其对类风湿关节炎的免疫抗炎作用,为开发紫草素成为COX-2抑制剂提供一定的实验基础。

Objective To investigate the effect of shikonin on the cyclooxygenase-2 （COX-2） mRNA expression in synovial fibroblasts and the immune anti-inflammatory mechanism of shiko- nin. Methods Isolated and cultured synovial membrane fibroblasts, added IL-I[3, TNF-a15 ng/mL respectively, human synovial cells of rheumatoid arthritis were simulated in vitro. As a positive control of celecoxib, synovial fibroblasts were interfered with shikonin in concentration of 5 -80 μg/L,then cultured for 24 h,48 h and 72 h. Semi-quantitative reverse transcriptionpolymerase chain reaction（RT-PCR） was used to detect the COX-2 mRNA expression of synovial fibroblasts. Results COX-2 mRNA expression of synovial fibroblasts was significantly dependent on medication time （ P 〈 0.0001 ）, not on the concentration （ P = 0. 089 ）. Celecoxib significantly inhibit the synovial fibroblasts and showed that the level of COX-2 mRNA expression without sinificant concentration-dependence （P = 0. 051 ）, only with medication time （P 〈 0. 0001）. Compared with celecoxib, the COX-2 mRNA expression effected by shikonin on synovial fibro-blasts had no significant difference in inhibititon. Conclusion Shikonin can inhibit COX-2 mRNA expression and reduce the production of inflammatory cytokines in order to play its immune anti-inflammatory role in rheumatoid arthritis. This study could provide some experimental basis for the development of shikonin as a COX-2 inhibitor.