大黄素对类风湿关节炎滑膜细胞增殖及转移的抑制作用

The inhibitory effects of emodin on the proliferation and metastasis of synovial cells form rheumatoid arthritis

目的探讨大黄素对体外培养类风湿关节炎患者成纤维样滑膜细胞增殖及转移的影响。方法采用组织块提取法和酶消化法体外培养类风湿关节炎滑膜细胞,分别加入不同浓度大黄素培养6、12、24 h,CCK-8法检测增殖能力的变化及时效关系,Transwell法检测处理前后转移能力的变化。结果酶消化法和组织块法分别于14 d和21 d得到原代RA成纤维样滑膜细胞,扩大培养3代后用大黄素处理,CCK-8检测6 h处理后20、40、60、80、100μmol/L大黄素的抑制率依次为0.39±0.11、0.51±0.04、0.55±0.07、0.72±0.18、0.98±0.11。且随着处理时间延长,抑制率增加(P<0.05)。Transwell法检测浓度为20、40、80μmol/L大黄素处理后,处理组穿过小室膜的细胞数明显少于对照组,迁移试验表明,抑制率分别为39.48%、54.76%、82.99%。侵袭试验表明,抑制率分别为25.38%、60.41%、81.73%。与对照组比较差异均具有统计学意义(P<0.05)。结论不同浓度的大黄素对体外培养RA-FLSs的转移及增殖能力具有抑制作用,且具有一定的量效和时效关系,为进一步研究大黄素在类风湿关节炎的治疗提供一定的实验室依据。

This study was designed to investigate the effects of emodin on the proliferation and metastasis of fibroblast-like synoviocytes （FLSs） from patients with rheumatoid arthritis （RA）. FLSs from rheumatoid arthritis were obtained by tissue block extraction method or enzyme digestion method, and then cultured in vitro. Different concentrations of emodin were used to incubate with FLSs for 6,12, or 24 hours, and then CCK-8 assay was used to test the effect of emodin on FLSs proliferation. Additionally, Fibroblast invasive ability was tested by transwell system. Data revealed that the original generation of RA-FLSs could be acquired by enzyme digestion method and tissue block extraction method in 14 d and 21 d, respectively. The results of CCK-8 displayed that the inhibiting rate of the different concentrations of 20, 40, 60, 80, 100 μmol/L emodin on FLSs proliferation were 0.39±0.11, 0.51±0.04, 0.55±0.07, 0.72±0.18, and 0.98±0.11 after 6 h incubation. Furthermore, as time went on, the inhibition got stronger （P〈0.05）. Migration test showed that the cells permeated the small room membrane was significantly less in 20, 40, and 80 μmol/L emodin treatment groups than that in control group, the inhibition rate were 39.48%, 54.76%, 82.99% respectively, while invasion test indicated the inhibition rate were respectively 25.38%, 60.41%, 81.73%, respectively. Taken together, emodin can obviously inhibit the proliferation, invasion and migration of RA-FLSs, which may provide laboratory basis for further research on the roles of emodin in the treatment of rheumatoid arthritis.