姜黄素治疗聚乙烯颗粒诱导骨溶解的体内实验
| 作者: |
1韩斐,
1胡懿郃,
1安森博,
1雷鹏飞,
1汪龙
1 中南大学湘雅医院骨科，长沙 410008 |
| 通讯: |
汪龙
Email: dr_wanglong@csu.edu.cn |
| DOI: | 10.3978/j.issn.2095-6959.2017.02.019 |
| 基金: | 国家自然科学基金, 81601883 湖南省卫生计生委科研基金, B2015-005 |
摘要
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(tartrate-resistant acid phosphatase，TRAP)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。
关键词：
骨溶解
无菌性松动
姜黄素
小鼠气囊植骨模型
NF-κB
c-Fos
NFATc1
NF-κB抑制蛋白
磷酸化NF-κB抑制蛋白
In vivo experiment of curcumin treating polyethylene particle induced osteolysis
CorrespondingAuthor: WANG Long Email: dr_wanglong@csu.edu.cn
DOI: 10.3978/j.issn.2095-6959.2017.02.019
Abstract
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.