去卵巢小鼠绝经后骨质疏松模型的建立和综合评定

Establishment and comprehensive evaluation of postmenopausal osteoporosis model in ovariectomized mice

目的建立和评定去卵巢小鼠骨质疏松模型,以期为后期进行不同的药物干预治疗提供准确的时机和模型。方法选取10~12周龄的C57BL/6雌性小鼠10只,随机分为假手术对照(Sham)组和卵巢摘除骨质疏松模型(OVX)组,每组5只。术后8周,将小鼠取血后处死,取一侧的股骨,剔除软组织,然后进行冰冻切片,另一侧进行显微断层扫描技术(Micro CT)检查。分别检测血钙、磷、镁水平,行组织病理学以及免疫荧光观察。结果血清学检测,OVX组与Sham组的钙、磷、镁含量变化不大,OVX组碱性磷酸酶含量比Sham组的含量低(P<0.01);Micro CT检查,OVX组小鼠骨小梁数量减少,间隔大,分离度也增加,排列稀疏,组织病理学HE染色发现OVX组破骨细胞数目增加(P<0.01);OVX组以及Sham组的碱性磷酸酶染色(ALP)的变化差异不大,抗酒石酸酸性磷酸酶(TRAP酶)表达的面积增加,颜色加深。结论C57BL/6雌性小鼠行双侧卵巢摘除法可以建立骨质疏松症模型,通过多种方法可以客观准确验证骨质疏松模型,以期为相关研究提供重要的参考依据。

Objective To establish and evaluate the osteoporosis model in ovariectomized mice,so as to provide appropriate timing and model for different drug interventions in later period.Methods Ten female C57BL/6 mice aged 10~12 weeks were selected and randomly divided into Sham operation control(Sham)group and ovariectomized osteoporosis model(OVX)group,with 5 mice in each group.At eight weeks after operation,the mice were sacrificed after blood collection.Their femur on one side was separated,soft tissue was removed,and then frozen section was made.The other side was examined by Micro CT.Lastly,their levels of serum calcium,phosphorus and magnesium were measured and observed by histopathology and immunofluorescence.Results Serological tests showed that the calcium,phosphorus and magnesium contents in OVX group and Sham group did not change significantly.The alkaline phosphatase content of OVX group was lower than that of Sham group(P<0.01).Micro CT examination showed that the number of bone trabeculae of rats decreased,the interval was large,the separation degree increased,and the arrangement was sparse in the OVX group.Histopathology HE staining showed increased number of osteoclasts in rats of OVX group(P<0.01).The change of alkaline phosphatase staining(ALP)in two groups showed little difference.Their expression area of trap-resistant acid phosphatase(TRAP)expanded,with color deepened.Conclusion Postmenopausal osteoporosis model can be established by bilateral ovariectomy in female C57BL/6 mice.And the osteoporosis model can be verified objectively and accurately by various methods,so as to provide an important reference for relevant studies.