淫羊藿苷对钛颗粒诱导的炎症性骨溶解的抑制作用  

Inhibitive effect of icariin on titanium particle-induced inflammatory osteolysis

作　　者：邵洪国[1] 崔京福 朱世军[1] 王熠军 朱锋[1] 杨惠林[1] 耿德春[1] 徐耀增[1] 

机构地区：[1]苏州大学附属第一医院骨科,215006

出　　处：《中华创伤杂志》2015年第9期850-855,共6页Chinese Journal of Trauma

基　　金：国家自然科学基金资助项目（81472077,81372018）

摘　　要：目的探讨淫羊藿苷对钛颗粒诱导炎症性骨溶解的治疗作用。方法雄性C57BL／J6小鼠84只，按随机数字表法分为空白组、对照组、淫羊藿苷低剂量治疗组（L组）和高剂量治疗组（H组），每组21只。治疗组采用钛颗粒诱导小鼠颅骨溶解模型，并给予不同浓度的淫羊藿苷：L组0．1mg·g-1·d-1，H组0．3mg·g-1·d-1，持续至建模后14d。采用显微CT评估骨溶解程度；HE染色观察细胞浸润程度；形态计量学测量骨溶解面积（EBS）；抗酒石酸酸性磷酸酶（TRAP）染色检测成熟破骨细胞；定量RT—PCR检测降钙素受体（CTR）、TRAP、组织蛋白酶K（Cath—K）和激活T细胞的核因子c1（NFATcl）基因mRNA含量；免疫组织化学检测肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL—1β）和IL-6的表达变化；酶联免疫吸附（ELISA）法检测核因子-kβ受体活化因子配体（RANKL）和骨保护素（OPG）的表达量。结果显微CT显示治疗组颅骨骨溶解现象明显减轻，骨密度、骨体积分数L组和H组分别为（89．44±18．51）mg／cm2、（48．32±9．54）％和（144．85±19．49）mg／cm2、（65．67±6．80）％，与对照组（61．23±13．41）mg／cm2、（27．83±7．51）％比较，差异有统计学意义（P〈0．05）。HE染色表明，对照组骨膜内有大量巨噬样细胞，L组和H组炎症反应明显减轻。对照组EBS为（0．14±0．03）mm2，L组和H组EBS降为（0．07±0．01）mm2和（0．04±O．01）mm2（P〈0．05）。对照组颅骨溶解侧可见大量TRAP阳性细胞，治疗后TRAP阳性细胞明显减少。免疫组织化学染色表明淫羊藿苷能抑制TNF-α、IL-1β和IL-6的表达。RT—PCR显示，钛颗粒上调CTR、TRAP、Cath—K和NFATcl基因mRNA表达，经治疗后上述基因mRNA表达水平较低，与对照组比较差异有统计学意义（P〈0．05）。ELISA法检测结果显示，对照组RANKL和OPG的表达量分别为（23．02±4．39）og�Objective To determine the therapeutic effect of icariin in the treatment of titanium particle-induced inflammatory osteolysis and examine the relationship between icariin and receptor activator of NF-KB ligand （ RANKL）/osteoprotegerin （OPG）. Methods Eighty-four male C57BL/J6 mice were di,lided into four groups （21 animals for each） according to the random number table： black group, control group （titanium particles only）, low-dosage icariin group （0. 1 mg g-1 . d-1, L group）, and high-dosage icariin group （0. 3 mg g-1 d-1 , H group）. The animal model of titanium particle- induced calvaria osteolysis was used in this study. After 2 weeks mice calvariae were harvested for micro- CT, histological and molecular analysis. Results As assessed by the micro-CT, icariin reduced parti- cle-induced osteolysis： bone mineral density and bone volume fraction in L group was （89. 44 ± 18.51 ） mg/cm2 and （48.32±9.54） % and was （ 144.85 ± 19.49 ） mg/ cm2 and （65.67±6.80） % in H group as compared to （61.23± 13.41 ）mg/cm2 and （27.83± 7.51 ）% in control group （P 〈 0.05 ）. HE staining demonstrated diminished inflammation after treatment with ieariin while intensive maerophage infiltration into the calvariae in control group. Histomorphometric results demonstrated the area of eroded bone surface （EBS） was reduced to （0.07 ± 0.01 ）mm2 and （0.04 ± 0.01 ）mm2 respectively in L and H groups compared to （0.14 ± 0.03）mm2 in control group （P 〈 0.05）. Tartrate resistant acid phosphatase （TRAP） staining revealed multiple TRAP-positive cells accumulated along the EBS in control group, whereas the number of TRAP-positive cells was reduced by icariin. Immunohistologieal examination showed titanium particle up-regulated expressions of tumor necrosis factor - α （TNF-α） , interleukin （IL）-1β and IL-6. In contrast, low expressions were noted after ieariin treatment. RT-PCR showed titanium particle stimulation could enhance mRNA expressions of eale

关 键 词：关节成形术 置换 骨质溶解 破骨细胞 钛 

分 类 号：R285.5[医药卫生—中药学]