姜黄素治疗聚乙烯颗粒诱导骨溶解的体内实验

In vivo experiment of curcumin treating polyethylene particle induced osteolysis

目的:探讨姜黄素对聚乙烯颗粒诱导骨溶解的治疗效果及作用机制,为防治假体无菌性松动提供新思路。方法:建立小鼠气囊植骨模型,随机分为:空白组、聚乙烯组、溶剂组、姜黄素组(姜黄素+溶剂)。干预完成后杀死小鼠,取出背部气囊组织及其内植骨骨片。行抗酒石酸酸性磷酸酶(tar trate-resistant acid phosphatase,TR AP)染色检测骨片中破骨细胞的数量。QPCR检测气囊组织中c-Fos,NFATc1的mRNA表达水平,Western印迹检测气囊组织中IκBα以及p-IκBα的蛋白含量。结果:TRAP染色发现,聚乙烯组和溶剂组骨片中染色的破骨细胞较多,而空白组和姜黄素组极少或没有染色的破骨细胞。QPCR检测基因表达情况:c-Fos,聚乙烯组>溶剂组>空白组>姜黄素组,前两组与后两组相比,组间差异有统计学意义(P<0.01);NFATc1,聚乙烯组>溶剂组>空白组>姜黄素组,前两组与后两组相比,组间差异有统计学意义(P<0.01)。Western印迹检测IκBα蛋白相对灰度值:姜黄素组>空白组>溶剂组>聚乙烯组,前两组与后两组相比,组间差异有统计学意义(P<0.01);p-IκBα蛋白相对灰度值:聚乙烯组>溶剂组>空白组>姜黄素组,前两组与后两组相比,组间差异有统计学意义(P<0.01)。结论:在小鼠气囊植骨模型中,姜黄素可以通过抑制NF-κB的活化以及c-Fos,NFATc1的表达来抑制超高分子量聚乙烯(ultra-high molecular weight polyethylene,UHMWPE)诱导的骨溶解反应。

Objective： To investigate the therapeutic effect and mechanism of curcumin on polyethylene particle induced osteolysis, and provide new insights into the prevention and treatment of prosthetic aseptic loosening. Methods： The mouse models of bone graft air pouch were established and randomly divided into the following four groups： blank group, polyethylene （PE） group, solvent group, curcumin group （curcumin ＋ solvent）. After the intervention, all the mice were harvested and the air pouchs and skulls were taken out. Tartrate-resistant acid phosphatase （TRAP） staining was used to detect the amount of osteoclast in skull. Q-PCR was performed to measure the mRNA level of c-Fos, NFATc1; Westen blot was performed to test the protein level of IκBα and p-IκBα in air pouch tissue. Results： The TRAP staining result showed that dyed osteoclast in polyethylene and solvent group were more than that in the other two groups; however, blank and curcumin group showed little or no osteoclast dyeing. QPCR： the mRNA level of c-Fos, polyethylene group 〉 solvent group 〉 blank group 〉 curcumin group, compared the former two groups with later two, the difference was statistically significant （P〈0.01）; the mRNA level of NFATc1, polyethylene group 〉 solvent group 〉 blank group 〉 curcumin group, compared the former two groups with later two, the difference was statistically significant （P〈0.01）. Western blot： the IκBα protein grey value, curcumin group 〉 blank group 〉 solvent group 〉 polyethylene group, compared the former two groups with later two, the difference was statistically significant （P〈0.01）; p-IκBα protein grey value, polyethylene group 〉 solvent group 〉 blank group 〉 curcumin group, compared the former two groups with later two, the difference was statistically significant （P〈0.01）. Conclusion： In air pouch bone graft model of mice, curcumin could reduce ultra-high molecular weight polyethylene induced bone dissolution reaction by inhibiting NF-κB activation and c-FOS, NFATc1 expression.