Periostin抗体的制备和Periostin组织表达特异性的研究

目的:制备抗Periostin抗体,并了解Periostin的组织表达特性。方法:以重组、纯化的小鼠Periostin为抗原,混入Freund完全佐剂充分乳化后,免疫新西兰大白兔;用双向免疫扩散试验检测抗体效价,将获得的多克隆抗血清初步纯化后,用Western blot和免疫组化检测小鼠Periostin的组织表达特异性。结果:Western blot显示Periostin在小鼠颅盖骨及牙周组织总蛋白中有特异性表达,而在肝、肾和脑组织总蛋白中没有表达。免疫组化检测,下颌骨表面骨膜和牙周膜组织中的Periostin染色呈强阳性,而在牙骨质、牙本质、牙槽骨和牙髓组织中则为阴性,并且它不是粘附于细胞表面,而是分布在由成骨细胞和牙周膜细胞分泌于其周围的细胞外基质中,牙槽骨中的骨细胞也不产生Periostin。结论:Periostin这种在有骨形成能力组织中的高表达,而在成熟骨组织中无表达的特点提示了其可能与矿化组织的早期形成有关。

Preparation of Periostin antibody and the expression of Periostin in mice

Objective:To prepare a rabbit anti-mouse Periostin IgG antibody and to study the expression of Periostin in mice.Methods:Complete Freund adjuvant with recombinant Periostin was used to immunize rabbits.Whole blood was collected from the animals after immunization for 5 times. Serum was obtained by centrifugation. The specific IgG antibody was purified from the serum by ammonium sulfate. Total protein isolated from mouse liver, brain, kidney and calvaria and periodontal tissue was used to study the expression of Periostin in mice by Western blot, immunohistochemical staining was applied to examine Periostin expression in jaw bone and dental tissues.Results:Anti-Periostin antibody with the titer of 128 was obtained.Western blot showed that there was a specific band near Mr 90 000 in calvaria and periodontal tissues, but not in liver, brain and kidney.By immunohistochemical examination Periostin was detected in both mandibular periosteum and periodontal ligament,but not in cementum, dentin, alveolar bone and dental pulp.Conclusion:Periostin may play a role in bone and periodontal tissue formation.