姜黄素对类风湿关节炎患者破骨细胞生成数量和活性的影响

Effects of curcumin on the number and activity of osteoclasts in patients with rheumatoid arthritis

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 investigate the effects of curcumin on the osteoclastogenesis from rheumatoid arthritis（RA）patients in vitro. Methods Peripheral blood mononuclear cells（PBMCs）from patients with RA separated by density centrifugation were cultured in the medium containing M-CSF and RANKL,added with different concentrations of curcumin in the process.There were 4 groups named blank control group,curcumin 2.5 μmol·L -1 group,curcumin 5 μmol·L -1 group and curcumin 10 μmol·L -1 group.After cultured for 14 days,TRAP staining was used for the detection and calculation of osteoclasts and the activity of TRAP was evaluated.Results After cultured for 14 days,the number of TRAP-positive osteoclasts were decreased significantly in curcumin 2.5 μmol·L -1 group （96.89 ±3.51）,curcumin 5 μmol·L -1 group（76.44 ±1.88）,curcumin 10 μmol·L -1 group（62.56 ±2.70）,which were signifi-cantly lower than that in the blank control group（131.00 ±4.03,P 〈0.05）.The activities of TRAP in 2.5 μmol·L -1 group,5 μmol ·L -1 group and 10 μmol·L -1 group were 5.74 ±0.36 U·L -1 ,4.21 ±0.12 U·L -1 and 3.06 ±0.07 U·L -1 ,which were lower than that in the blank control group（7.48 ±0.22 U·L -1 ,P 〈0.05）.Conclusions Curcumin can inhibit the osteoclastogenesis of the PBMCs from RA patients,and the inhibition effect is enhanced with the increase in the concentration of curcumin.