转染hIGF-1基因增强软骨细胞聚集蛋白多糖、Ⅱ型胶原的表达

Influence of hIGF-1 gene on chondrocytes aggrecan,collagen typeⅡ expression

目的探讨人胰岛素样生长因子(human insulin-like growth factor,hIGF-1)基因对软骨细胞分泌聚集蛋白多糖(aggrecan)、Ⅱ型胶原的影响.方法构建并鉴定携带hIGF-1基因的重组腺病毒(Ad/CMV-hIGF-1),采用1、10、100及500感染复数单位(multiplicity of infection,MOI)的Ad/CMV-hIGF-1转染软骨细胞,PBS为阴性对照,100μg/L hIGF-1生长因子为阳性对照,转染后4 d进行aggrecan、Ⅱ型胶原免疫组化,参照文献分析免疫组化阳性单位.结果在1～100MOI范围内,随着Ad/CMV-hIGF-1滴度增加,aggercan、Ⅱ型胶原表达递增,500 MOI Ad/CMV-hIGF-1转染后,aggrecan表达骤减;Ⅱ型胶原表达下降不明显.结论 hIGF-1基因对软骨细胞aggrecan、Ⅱ型胶原的表达有影响;100 MOI Ad/CMV-hIGF-1优于100 μg/L hIGF-1生长因子的作用.

ObjectiveTo study the role of human insulin-like growth factor-1 gene on aggrecan, collagen type expression of human articular chondrocytes. MethodsRecombination replication defective adenovirus with hIGF-1 gene (Ad/CMV-hIGF-1) was constructed and identified. Human embryo articular chondrocytes were infected with 1, 10, 100, or 500 MOI Ad/CMV-hIGF-1 with PBS as negative control and hIGF-1 growth factor as positive control. Aggrecan, or collagen type immunohistochemistry was taken 4 days after infection. Immunohistochemistry positive units were analyzed according to the reference reported. ResultsAggrecan, or collagen type expression increased as Ad/CMV-hIGF-1 increasing at the range of 1 to 100 MOI, but at the point of 500 MOI, aggrecan expression abruptly decreased, collagen type expression was the same as that of the 100 MOI. Aggrecan, or collagen type expression of chondrocytes infected with 100 MOI Ad/CMV-hIGF-1 was about two to three times as that of the PBS group; It was higher than that of 100 μg/L hIGF-1 growth factor. ConclusionThe results indicate that hIGF-1 gene has influence on chondrocytes aggrecan, or collagen type expression.