原位杂交法检测骨和软骨组织中Ⅰ、Ⅱ型胶原基因的表达  

Detection of Type Ⅰand Type Ⅱ Collagen mRNA in Bone and Cartilage by in Situ Hybridization

作　　者：陈克明[1] 刘兴炎[1] 葛宝丰[1] 王勇[1] 白孟海[1] 

机构地区：[1]兰州军区总医院骨科研究所,兰州市730050

出　　处：《骨与关节损伤杂志》2000年第5期359-360,共2页The Journal of Bone and Joint Injury

摘　　要：目的　用原位杂交法研究骨和软骨组织中Ⅰ、Ⅱ型胶原基因的表达。方法　以与Ⅰ型、ⅡA型和ⅡB型胶原mRNA特异性互补的寡核苷酸序列为探针 ,用末端转移酶将地高辛标记于 3′ -末端 ,在兔胫骨近端组织切片上进行Northen原位杂交 ,检测关节软骨、骺板和新生骨小梁区三种胶原基因的表达情况。结果　关节软骨和骺板区均检测到软骨细胞中有ⅡB型胶原基因表达 ,新生骨小梁区检测到成骨细胞中有Ⅰ型胶原基因表达。结论　本实验方法灵敏度高 ,结果可靠 。Objective To investigate gene expression of type Ⅰ and type Ⅱ collagen in bone and cartilage by in situ hybridization.Methods Three oligonucleotide probes which were specific seperately for type Ⅰ,type Ⅱ A and type ⅡB collagen were sythesized with DNA synthesizer,and labelled at 3'-end by DIG-ddUTP.The specimens of normal rabbit proximall tibia including articular cartilage,epiphysis and new-produced trabecula were cut into 5um sections and hybridized with the oligonucleotide probes.Results Type ⅡB collagen mRNA was clearly demonstrated in chondrocytes in articular cartilage and epiphysis,type Ⅰ collagen mRNA was localized in osteoblasts lining in surface of newproduced trabecula.Conclusions Our method of in situ hybridization is comparatively simple and safe,which can be used to further studies on relationship between collagen gene expression and bone disease and osteogenesis.

关 键 词：原位交 骨组织 软骨组织 Ⅰ胶原 基因表达 

分 类 号：R681.04[医药卫生—骨科学]