摘要
目的 研究转化生长因子 - β1 (TGF- β1 )基因转染鼠成骨细胞对其生物学特性的影响。方法 将TGF- β1 基因导入成骨细胞 ,通过原位杂交检测和成骨细胞培养上清液 TGF- β1 活性测定 (水貂肺上皮细胞生长抑制试验 ) ,了解转染细胞 TGF- β1 的表达。检测基因转染及转染细胞上清对成骨细胞增殖和碱性磷酸酶 (AL P)活性的影响 ,观察 TGF- β1 基因转染成骨细胞生物学活性的变化。结果 原位杂交检测基因转染成骨细胞可明显表达TGF- β1 。上清液 TGF- β1 活性检测结果显示 1∶ 1、1∶ 2、1∶ 4复合培养基对 Mv1抑制率分别为 16 .3%、2 2 .7%和2 8.2 % ,上清中 TGF- β1 含量越高 ,活化后对 Mv1抑制作用越明显。 TGF- β1 基因转染对成骨细胞生物学活性无明显影响 ,而转染细胞上清活化后可促进成骨细胞增殖 ,抑制 AL P活性。结论 TGF- β1 基因转染鼠成骨细胞可显著促进 TGF- β1 表达 ,转染细胞生物学特性保持稳定 。
Objective To investigate the effect of transforming growth factor β 1 (TGF β 1) gene transfer on the biological characteristics of osteoblasts. Methods The expression of TGF β 1 in the transfected osteoblasts was detected by in situ hybridization and assay of TGF β 1 activity in the supernatant (mink lung epithelium cell growth inhibition test). The effects of gene transfer and supernatant of the transfected osteoblasts on the proliferation and alkaline phosphatase(ALP) activity of osteoblasts were detected by 3H TdR and MTT. Results The results of in situ hybridization analysis suggested that the osteoblasts transfected by TGF β 1 gene could express TGF β 1 obviously. The complex medium, which was the mixture of serum free DMEM and the activated supernatant according to 1∶1, 1∶2, 1∶4, could inhibit growth of Mv 1 Lu evidently and the ratios of inhibition were 16 3%, 22 7%, 28 2% respectively. TGF β 1 gene transfer had no effect on the biological characteristics of osteoblasts, but the activated supernatant of transfected osteoblasts stimulated proliferation and inhibited ALP activity of osteoblasts. Conclusion TGF β 1 gene transfer promotes the expression of TGF β 1 and the biological characteristics of transfected osteoblasts are stable, which is helpful for gene therapy of bone defects in vivo.
出处
《中国修复重建外科杂志》
CAS
CSCD
2002年第2期86-88,共3页
Chinese Journal of Reparative and Reconstructive Surgery
