淫羊藿苷口服给药对骨质疏松牙周炎小鼠牙槽骨吸收的影响

Effect of oral administration of icariin on periodontitis associated alveolar bone resorption in an osteoporosis mouse model

目的 :研究口服淫羊藿苷(ICA)对骨质疏松小鼠牙周炎引起的牙槽骨吸收的抑制作用。方法 :3月龄、雌性、C57BL/6J小鼠随机分为3组,即正常组(SHAM组)、卵巢切除+口腔涂布牙龈卟啉单胞菌(Pg)+淫羊藿苷组(OVX+Pg+ICA)、卵巢切除+Pg口腔涂布组(OVX+Pg)。小鼠适应性喂养1周,第2周进行双侧卵巢切除术,诱导小鼠骨质疏松形成。从第4周开始,对小鼠牙周涂布Pg,1次/d,连续1周。第12周收集左侧下颌骨进行固定切片及染色,分析各组之间牙槽骨吸收高度的差异。收集右侧下颌骨进行亚甲蓝染色,分析各组间牙槽骨吸收面积的差异;收集双侧上颌骨牙周组织蛋白,分析成骨相关蛋白的表达差异。采用SPSS16.0软件包对数据进行统计学分析。结果:小鼠股骨及牙周组织切片染色显示,成功建立了小鼠骨质疏松牙周炎模型。对牙槽骨吸收距离和面积的测量分析显示,相比于OVX+Pg组,口服淫羊藿苷可显著减少釉-牙骨质界-牙槽嵴顶(CEJ-ABC)的距离及颊舌侧牙槽骨吸收面积(P<0.05);Western免疫印迹显示,相比于OVX+Pg组,OVX+Pg+ICA组中Runx2、OSX、OCN及OPN蛋白表达水平显著升高(P<0.05)。结论:口服淫羊藿苷在预防小鼠骨质疏松发生的同时,可有效减少牙周炎引起的牙槽骨吸收。

PURPOSE: To study the effect of oral administration of icariin(ICA) on periodontitis associated alveolar bone resorption in an osteoporosis mouse model. METHODS: Three-month old, female C57 BL/6 J mice were randomly divided into 3 groups: normal group(SHAM), ovariectomy + oral smearing of Porphyromonas gingivalis(Pg)+ icariin group(OVX+Pg+ICA), ovariectomy+ Pg group(OVX+Pg). On the second week, the mice underwent ovariectomy to induce osteoporosis.On the fourth week, the mice were treated with Pg by oral smearing for 1 week, 1 time/day. The samples were harvested on the 12 th week: the left mandibles were harvested to make the sections and histological staining, to analyze the differences of bone resorption areas among the three groups;the right mandibles were harvested for methylene blue staining, to analyze the differences of bone resorption depth among the three groups. Protein from the periodontal tissues were extracted to analyze the differences of osteoblastic related protein expression level among the three groups. SPSS 16.0 software package was used for statistical analysis. RESULTS: Histological analysis of murine femurs and periodontal tissues demonstrated that the model of periodontitis in osteoporosis mice was successfully created in this study. Quantification of the histological staining showed that compared with OVX+Pg group, icariin treatment decreased the depth of CEJ-ABC and the areas of alveolar bone resorption(P<0.05);Western blot results showed icariin treatment could significantly increase the protein expression level of Runx2, OSX, OCN and OPN in periodontal tissues(P<0.05). CONCLUSIONS: Icariin treatment can effectively prevent both bone loss in osteoporosis and alveolar bone resorption induced by periodontitis.