摘要
目的观察选择性环氧化酶-2（cyclooxygenase-2，COX-2）抑制剂——帕瑞昔布对钛颗粒诱导骨溶解的治疗作用以及COX-2与NF—κB受体活化因子配体（receptor activator of NF—κB ligand，RANKL）／NF～KB受体（receptor activator of NF—κB，RANK）通路的关系。方法雌性BALB／c小鼠45只，其中15只作为颅骨供体，其余采用随机数字表法分为空白组、对照组和药物组，每组10只。采用钛颗粒诱导小鼠骨溶解模型，药物组建模前2d注射帕瑞昔布（6mg·kg^-1～·d^-1），持续至建模后2周。采用抗酒石酸酸性磷酸酶（tartrate—resistant acid phosphatase staining，TRAP）染色检测成熟破骨细胞；免疫组织化学检测COX-2和RANKL表达变化；ELISA法检测囊壁中前列腺素E2（dinoprostone，PGE2）、IL-1β和TNF—α含量；定量RT—PCR检测COX～2、IL-1β、TNF—α RANK和RANKL基因mRNA含量。结果TRAP染色结果显示，药物组TRAP阳性区域面积占植入骨片面积比例为（0．27±0．03）％，与对照组[（0．60±0．06）％]比较差异有统计学意义（P〈0．01）。免疫组织化学结果显示，钛颗粒植入后囊壁内COX-2和RANKL深染区域增多；药物组RANKL阳性区域减少，但COX-2表达未见明显减少。RT—PCR结果显示，钛颗粒上调COX-2、IL-1β、TNF-α、RANK和RANKL基因mRNA表达水平，药物组RANK和RANKL基因mRNA水平较低，与对照组比较差异有统计学意义（P〈0．01）。ELISA结果显示，药物组PGE2含量为（202．62±11．49）pg／ml，与对照组[（407．60±13．55）pg／m1]比较差异有统计学意义（P〈0．01）；药物组IL-1β和TNF-α含量与对照组比较差异无统计学意义（P〉0．05）。结论选择性COX-2抑制剂——帕瑞昔布能下调RANKL／RANK的表达，减少TRAP阳性破骨细胞数量，减轻骨溶解；但对COX-2以及TNF—Or．和IL-1β的表达未产生明显影响。
Objective To observe the effect of selective cyclooxygenase-2 （COX-2） inhibitor parecoxib in treatment of titanium-particle-induced inflammatory osteolysis and examine the relationship between COX-2 and receptor activator of NF-KB ligand （ RANKL）/receptor activator of NF-KB （RANK） signaling pathway. Methods Fifteen out of 45 female BALB/e mice were served as skull donators. The rest were divided into blank group, control group, and treatment group according to random number table, with 10 mice per group. Models of osteolysis in mice iuduced by titanium particles were estaelished. Treatment group were given parecoxib （6 mg· kg^-1· d ^-1） at 2 days prior to the modeling, which was continued tilled two weeks after the modeling. Mature osteoelasts were identified using tartrate-resistant acid phosphatase （TRAP） staining. Expressions of COX-2 and RANKL were detected using immunohistochemieal staining. Levels of dinoprostone （PGE2） , IL-β, and TNF-a in capsule wall wereexamined by ELISA method. Expressions of the mRNA for COX-2, IL-β, TNF-α, RANK and RANKL were determined by quantitative RT-PCR. Results TRAP staining revealed ratio of TRAP positive area to bone graft area was （0.27±0.03 ） % in treatment group, with statistical difference from control group [ （0.60±0.06） % ] （P 〈 0.01 ）. Immunohistochemical staining showed COX-2 and RANKL intensive area in capsule wall were increased after titanium particle stimulation. Instead, RANKL positive area was decreased in treatment group, but the decrease of COX-2 was insignificant. RT-PCR showed titanium particles stimulation could enhance the mRNA expressions of COX-2, IL-β, TNF-α, RANKL, and RANK. Whereas, mRNA expressions of RANK and RANKL in treatment group were reduced, with statistical difference in contrast to control group （P 〈 0. 01 ）. ELISA showed PGE2 level was （202.62± 11.49） pg/ml in treatment group, with statistical difference from control group [ （407.60± 13.55） pg/ml] （P 〈 0.01 ）. Levels of IL-β and TNF-α had no statistical differences between treatment group and control group （P 〉 0.05 ）. Conclusions Selective COX-2 inhibitor parecoxib impedes expression of RANK/RANKL, reduces the number of TRAP positive osteoclasts, and thereafter ameliorates osteolysis. Whereas, parecoxib presents no significant influence on expressions of COX2, TNF-α, and IL-β.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2013年第6期545-549,共5页
Chinese Journal of Trauma
基金
国家自然科学基金资助项目（81272018,81101399）
汀苏省自然科学基金资助项目（BK2011303）
江苏省卫生厅科研资助项目（H201012）
江苏省高校自然基金资助项目（10KJB320019）
苏州大学优秀博士选题重点资助项目
