蛇床子素对TCP颗粒诱导小鼠颅骨溶解的影响

 目的: 观察蛇床子素对磷酸三钙(tricalcium phosphate,TCP)颗粒诱导小鼠颅骨溶解的影响。方法: 采用TCP颗粒诱导小鼠颅骨溶解模型,于术后第2天颅顶局部注射蛇床子素(osthole)20 mg/kg,每周3次,持续干预2周。干预结束后处死动物取材,应用HE染色和抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRACP)染色观察假体周围破骨细胞生成和骨溶解程度;ELISA法检测血清和骨组织中骨转换标志物骨钙素(osteocalcin)水平、碱性磷酸酶(alkaline phosphatase,ALP)和TRACP活性以及骨膜中肿瘤坏死因子α (tumor necrosis factor-α,TNF-α)、白细胞介素6(interleukin-6,IL-6)和白细胞介素1β(interleukin-1β,IL-1β)水平。Western blot法检测颅骨组织葡萄糖调节蛋白 78(glucose-regulated protein 78,GRP78)和CAAT/增强子结合蛋白同源蛋白(CAAT/ enhancer binding protein homologous protein,CHOP)的表达变化。结果: 蛇床子素可明显抑制TCP颗粒诱导的破骨细胞生成,减少骨溶解面积,显著增加ALP和osteoclacin水平,降低TRAP活性并阻断炎症因子TNF-α、IL-6和IL-1β释放。此外,蛇床子素干预能明显减弱TCP颗粒激活的内质网应激反应。结论: 蛇床子素可抑制TCP颗粒诱导的小鼠颅骨溶解,其机制可能与抑制TCP颗粒诱导的内质网应激反应有关。

Effect of osthole on tricalcium phosphate particles-induced calvarial osteolysis in a mouse model

AIM: To observe the effect of osthole on tricalcium phosphate (TCP) particles-induced calvarial osteolysis in vivo.METHODS: Male ICR mice were randomly divided into sham group, TCP group and osthole group. A mouse calvarial model of osteolysis was established by TCP particles. On the second postoperative day, osthole (20 mg/kg) was locally injected into the calvarium under the periosteum 3 times a week. Two weeks after osthole treatment, blood and calvaria were collected to determine the level of bone turnover markers such as alkaline phosphatase(ALP), osteocalcin and tartrate-resistant acid phosphatase (TRACP). The periosteum was performed to examine the release of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β by ELISA. The calvaria was obtained for histological and molecular analyses. RESULTS: Data from HE and TRACP staining revealed that osthole prevented TCP particles-induced obvious increase in osteoclastogenesis and resorption area in the metaphysis of mouse calvaria. Osthole treatment increased ALP activity and osteocalcin level, and dncreased the activity of TRACP in the mouse serum compared with TCP group. Furthermore, TCP particles-induced the releases of TNF-α, IL-6 and IL-1β were significantly suppressed by osthole treatment. In addition, Western blot demonstrated that endoplasmic reticulum (ER) stress markers such as glucose-regulated protein 78 (GRP78) and CAAT/enhancer binding protein homologous protein (CHOP) were significantly up-regulated in TCP particles-implanted calvarial mice, indicating that TCP particles triggered an ER stress response in the mouse calvarial osteolysis model, which obviously attenuated by osthole. CONCLUSION: Osthole inhibits TCP particles-induced calvarial osteolysis in mice, which is mediated by inhibition of ER stress signaling pathway.