姜黄素调节核转录因子-κB信号通道抑制磨损颗粒诱导骨溶解

The study of curcumin modulates nuclear factor-kappa B signaling pathway to inhibit wear particle inducted osteolysis in vitro

目的建立颗粒诱导骨溶解体外模型，探讨姜黄素调节核转录因子-κB（NF-κB）信号通道对骨溶解的作用。方法将超高相对分子质量聚乙烯（UHMWPE）和小鼠单核/巨噬细胞株RAW264.7细胞共培养，构建磨损颗粒诱导模型，加入姜黄素干预，分为4组：A：空白组；B：聚乙烯颗粒诱导组；C：聚乙烯颗粒诱导＋低浓度姜黄素处理组；D：聚乙烯颗粒诱导＋高浓度姜黄素处理组。提取每组细胞的蛋白，用Western blot检测NF-κB通路中的关键蛋白核转录因子抑制蛋白（IκBα）、磷酸化核转录因子抑制蛋白（p-IκBα）的表达。相同的分组方法处理细胞后，提取每组细胞的总RNA，使用实时定量聚合酶链反应（Real-time PCR）法检测NF-κB通路中转录因子c-Fos和活化T细胞核因子1蛋白（NFATc1）的表达。结果（1）Western blot实验结果，B组IκBα与内参β-肌动蛋白（β-actin）灰度值之比IκBα/β-actin（0.029±0.004）低于其余3组（P=0.001）（A组：0.321±0.014，C组：0.352±0.019，D组：0.554±0.009），C组与D组比较（P=0.001）；B组磷酸化IκBα/β-actin（1.450±0.081）高于其余3组（P=0.001）（A组：0.593±0.041，C组：0.480±0.023，D组：0.511±0.024），C组与D组比较差异无统计学意义（P=0.163）。（2）Real-time PCR实验结果，B组中c-Fos （1.243±0.040）和NFATc1（1.255±0.009）的表达量最高（P=0.000），C组中c-Fos（0.868±0.032）高于D组c-Fos（0.814±0.009，P=0.015），C组中NFATc1（0.956±0.016）高于D组中NFATc1（0.624±0.009，P=0.001）。结论姜黄素可通过抑制NF-κB通道的激活抑制假体周围骨溶解的发生。

ObjectiveTo investigate the effect of curcumin on osteolysis through regulating nuclear factor-kappa B （NF-κB） signal pathway, by establishing the model of wear particle induced osteolysis in vitro.MethodsWear particles induced cellular model of osteolysis was build by co-culturing ultra high molecular weight polyethylene （UHMWPE） with mouse monocytes/macrophages RAW264.7 cells, intervented by adding curcumin. Four groups were involved： A： blank group; B： polyethylene particle induced group; C： polyethylene particle induced ＋ low concentration of curcumin treatment group; D： poly-ethylene particle induced ＋ high concentration of curcumin. Proteins were extracted from each group to detect the expression of the key protein inhibitor of NF-κB （IκB）and phosphorylated IκB （p-IκB） in the NF-κB pathway by using Western blotting. The same grouping method was used to detect the expression of transcription factor c-Fos and nuclear factor of activated T-cells cytoplasmic 1 （NFATc1） by using real time polymerase chain reaction （Realtime-PCR）.Results（1） Western blotting results： in group B, the grave value ratio of IκBα and β-actin （IκBα/β-actin） was 0.029±0.004, which was lower than the other three groups （P=0.001） （group A： 0.321±0.014, group C： 0.352±0.019, group D： 0.554±0.009）. Comparing group C with group D, the P value was 0.001; Group B’s p-IκBα/β-actin was 1.450±0.081, which was higher than other three groups （P=0.001） （group A： 0.593±0.041, group C： 0.480±0.023, group D： 0.511±0.024）. The comparison between group C and group D had no statistical significance （P=0.163）. （2） Real-time PCR results： expression of c-Fos （1.243±0.040） and NFATc1 （1.255±0.009） was the highest. Group C’s c-Fos expression （0.868±0.032） was higher than group D’s （0.814±0.009） （P=0.015）. Group C’s NFATc1 expression （0.956±0.016） was higher than group D’s （0.624±0.009） （P=0.001）.ConclusionCurcumin can inhibit the occurrence of periprosthetic osteolysis by inhibiting the activation of NF-κB signal pathway in vitro.