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目的研究姜黄素对类风湿关节炎(RA)患者破骨细胞生成数量和活性的影响。方法采集RA患者外周血,密度梯度离心法分离外周血单个核细胞(PBMCs),经核因子κB受体活化因子配体(RANKL)和巨噬细胞集落刺激因子(M-CSF)诱导分化,分别采用不同浓度(2.5、5、10μmol·L~(-1))姜黄素进行干预;根据姜黄素浓度将实验分为4组,即空白对照组、姜黄素低浓度组、姜黄素中浓度组和姜黄素高浓度组;14 d后行抗酒石酸酸性磷酸酶(TRAP)染色检测破骨细胞并计数;检测细胞TRAP活性。结果细胞培养14 d后,姜黄素低、中、高浓度组的TRAP阳性细胞计数(个/10个视野)分别为96.89±3.51、76.44±1.88和62.56±2.70,均低于空白对照组131.00±4.03(P<0.05);各浓度组的TRAP活性(U·L-1)分别为5.74±0.36、4.21±0.12和3.06±0.07,均低于空白对照组7.48±0.22(P<0.05)。结论姜黄素抑制RA患者PBMCs生成破骨细胞的数量和活性,且随着姜黄素浓度的增加,抑制作用呈增强趋势。
Objective To study the effect of curcumin on the number and activity of osteoclasts in patients with rheumatoid arthritis (RA). Methods The peripheral blood mononuclear cells (PBMCs) from patients with RA were collected and peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation and induced by RANKL and M-CSF (2.5, 5, 10μmol·L -1) curcumin were used for intervention. According to the concentration of curcumin, the experiment was divided into 4 groups: blank control group, curcumin low concentration group, curcumin middle concentration group and Curcumin high concentration group. After 14 days, TRAP staining was performed to detect osteoclasts, and the activity of TRAP was detected. Results After 14 days of cell culture, the TRAP-positive cell counts (a / 10 fields) of curcumin in low, middle and high concentration groups were 96.89 ± 3.51,76.44 ± 1.88 and 62.56 ± 2.70, respectively, which were lower than those in the blank control group 131.00 ± 4.03 (P <0.05). The TRAP activity (U · L-1) in each concentration group was 5.74 ± 0.36, 4.21 ± 0.12 and 3.06 ± 0.07, respectively, which were all lower than those in the control group (7.48 ± 0.22, P <0.05). Conclusion Curcumin inhibits the number and activity of osteoclasts produced by PBMCs in RA patients. With the increase of curcumin concentration, the inhibitory effect is enhanced.