石蜡包埋骨组织切片核酸原位杂交研究

Study of in situ hybridization in paraffin-embeded bone tissue slides

目的 探讨石蜡包埋骨组织切片的核酸原位杂交方法,以检测下颌骨骨折愈合过程中骨组织内整合素β_1mRNA的表达。方法 用地高辛标记的整合素β_1mRNA的寡核苷酸探针,对36只新西兰大耳白兔下颌骨骨折后不同愈合时期的骨组织标本石蜡切片进行核酸原位杂交,观察整合素β_1mRNA的时空变化规律。结果 在骨折愈合过程中的不同时期,整合素β_1mRNA在骨组织中的各种细胞的细胞质与细胞核中均呈阳性表达,骨折7天后整合素β_1mRNA表达增强,14～30天最强,90天后接近正常。结论 应用寡核苷酸探针进行核酸杂交技术,对于石蜡包埋骨组织的mRNA进行研究是可行的。

Objective To investgate the method of in-situ-hybridization using paraffin-embeded bone tissue slides for the detection of integrin?1 mRNA expression during mandibular fracture healing. Methods Using 36 NewZealand rabbits, mandibular fracture was created, in situ hybridization with Dig-lablled integr?1, mRNA oligonucleotide probe was performed. In different stage of fracture healing, the staining pattern both spatially and time - dependently was observed. Results Integrin?1 was widely espressed in fractured bone cells, the positive hybrid signal was found in the cytoplasm and nucleolus: After 7 days of fracture, increaseded expression of integrin?1 was observed, and it was peaked in 14 to 30 days, nearlly normal at 90 days. Conclusion By using oligonucleotide probe, it is feasible to performe the in situ hybridization study on paraffin-embeded bone tissue.