碱性成纤维细胞生长因子转染兔关节软骨细胞的研究

Enhancement of proliferation of rabbit chondrocytes transfected w ith basic fibroblast growth factor gene in vitro

目的 探讨碱性成纤维细胞生长因子 (bFGF)基因转染兔关节软骨细胞后对培养的关节软骨细胞形态、分裂增殖及代谢等方面的影响。方法 将bFGF基因克隆于真核表达载体pHβ。AP 1中 ,构建重组真核表达载体 pHβ bFGF ,转染兔关节软骨细胞。G418筛选阳性克隆 ,检测阳性细胞bFGF基因的表达水平。测定培养软骨细胞的DNA含量、糖醛酸含量、软骨细胞增殖情况及进行细胞周期分析。结果 bFGF基因转染软骨细胞表型未见显著变化 ;bFGF基因转染组、载体对照组、空白对照组DNA含量分别为 ( 77.37± 6 .2 1)、( 40 .39± 4.33)、( 33 .77± 4.2 5 ) μg/瓶 (P <0 .0 1) ,糖醛酸含量分别为 ( 30 8.8± 10 .2 )、( 77.9± 8.7)、( 80 .2± 10 .5 ) μg/瓶 ( P <0 .0 1) ,软骨细胞G1期分别为 5 9.3± 2 .1、6 9.5± 4.0、73 .1± 3 .9(P <0 .0 5 )。结论 bFGF转染关节软骨细胞后 ,可显著促进细胞分裂增殖并缩短细胞周期 ,为软骨组织工程研究提供新的技术路线及理论基础。

Objective To explore the proliferation ch aracteristics of chondrocytes transfected with pHβ-basic fibroblast growth fac tor (bFGF) expression vector.Methods The recombined expre ssion vector pHβ-bFGF was constructed and transfected into rabbit chondrocytes by lipofectamine.The phenotype changes of cells were observed under light micr oscopy and the DNA content,glucuronic acid,proliferation of cells and cell lif e phases were measured.Results The phenotypes of chondroc ytes were verified significantly except the bFGF transfected group.As compared with vector and blank group, the DNA content,glucuronic acid and proliferation of cells were dramatically increased and the G 1 phase of cell life cycle was shortened in the bFGF transfected group (P<0.01).Conclusion The transfection of bFGF gene into rabbit chondrocytes can shorten the cell cycle and enhance the cell proliferation,which may provide some experiment al data for cartilage histoengineering.