白黎芦醇对小鼠磨损颗粒诱导炎性骨溶解的抑制作用

The Influence of Resveratrol Inhibit Wear Debris-induced Inflammatory Osteolysis in a Murine Osteolysis Model

目的观察白藜芦醇对小鼠磨损颗粒诱导炎性骨溶解的抑制作用。方法 70只雄性BALB/c小鼠,其中45只小鼠随机分为对照组、模型组和药物组,每组15只。第1天,3组小鼠行腹腔注射麻醉,在小鼠背部皮下注射2mL空气,第2～6天再向气囊中注射0.5mL空气,形成气囊;另外25只小鼠作为供体,过量麻醉处死后取出颅骨。将受体小鼠麻醉后,背部气囊上做一个5 mm长的切口将颅骨通过切口植入气囊内。24h后,对3组小鼠作如下处理,对照组:向小鼠气囊内注射0.3mL无菌PBS液;模型组:用lmL无菌注射器向小鼠气囊内注射0.3mL处理好的钛颗粒,以激发炎症反应和溶骨反应;药物组:建模当日经灌胃予小鼠白藜芦醇[600mg·(g·d)^(-1)]。每天灌胃1次,持续14d,钛颗粒注射同模型组。3组小鼠于14d后行过量麻醉处死取材。结果 HE染色结果显示模型组植入气囊的颅骨骨膜出现骨溶解表现,并出现炎性反应;3组小鼠气囊囊壁厚度及炎性细胞总数差异均有统计学意义(P均<0.05),模型组小鼠的气囊囊壁厚度、囊壁炎性细胞计数高于药物组和对照组(P<0.05)。TRAP染色结果显示对照组可见零星散布的点状阳性紫红色的破骨细胞;模型组颅骨骨块可见大片紫红色区域,说明此区域的破骨细胞增殖活跃。药物组可见部分区域有紫红色的破骨细胞出现;ELISA检测结果显示药物组TNF-α、MMP-9及IL-6的表达量均低于模型组(P均<0.05)。结论白藜芦醇可能通过降低TNF-α、MMP-9及IL-6的表达量减轻小鼠磨损颗粒诱导的炎症骨溶解反应。

Objective To explore the influence of resveratrol inhibit wear debris-induced inflammatory osteolysis in a murine osteolysis model. Methods A total of 70 male BALB/c mice were used in the model, 45 mice out of them were randomly divided into control group, model group and titanium particles-stimulated group, with 15 mice in each group. On the first day, the three groups of mice were anesthetized by intraperitoneal injection, and at first beginning 2mL air was injected, 0.5mL air was injected subcutaneously from next day to sixth day into the back of the mice to form an air bag; the other 25 mice were used as donor, and the skull was removed after excessive anesthesia. After the recipient mice were anesthetized, a 5mm long incision was made on the back airbag to insert the skull through the incision into the balloon. After 24h, the three groups of mice were treated as follows： the control group： injection of 0.3mL aseptic PBS solution to the mouse air bag; model group： injection of lmL sterile syringe to the mice airbag injected with good titanium particles, to stimulate the inflammatory reaction and bone dissolution reaction; drug group： the drug group was injected stomach to the mouse resveratrol [ 600mg· （g· d）- 1 on the day of modeling. Once a day, continuous for 14d, titanium particles injection with the model group. The mice in the three groups were sacrificed under excessive anesthesia after 14 days. Results The results of HE staining showed that the periosteum of the skull periosteum appeared in the model group and appeared inflammatory reaction. The thickness of balloon wall and the total number of inflammatory cells in the three groups were statistically significant（P〈0.05）. The thickness of balloon wall and the count of inflammatory cells in the capsule wall were higher in the model group than in the drug group and the control group （P〈0.05）. The results of TRAP staining showed that there were sporadic scattered point positive purple red osteoclasts in the control group, and a large area of purple red in the skull bone of the model group, indicating that the proliferation of osteoclast in this area was active. In the drug group, the presence of upura osteoclasts in some areas showed that the expression of TNF-α, MMP-9 and IL-6 in the drug group was lower than that in the model group（P〈0.05）. Conclusion Resveratrol may reduce the wear particles induced inflammatory osteolysis by reducing the expression of TNF-α, MMP-9 and IL-6.