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目的 探讨转化生长因子 β1(TGF β1)基因转染骨髓基质细胞 (BMSCs)的表达能力。方法 取 6周龄新西兰白兔BMSCs ,传代培养后以 3μl阳离子脂质体 (Lipofectamine)∶1μgpcD NA3 TGF β1的比例转染 ,通过免疫组织化学染色及图像分析对 3组 15份样本检测TGF β1表达水平 ;利用水貂肺上皮细胞生长抑制法检测TGF β1生物活性 ,分析转染的量效关系和时效关系。结果 瞬时表达组转染后 48h免疫组织化学染色部分细胞呈强阳性 ,稳定表达组全部细胞呈强阳性且表达持续 4周以上。图像分析显示转染细胞TGF β1表达显著增强 ,与对照组比较差异有非常显著性 (P <0 .0 1)。量效关系显示 ,在转染体系中pcDNA3 TGF β1( μg)∶阳离子脂质体 ( μl)为1∶3时 ,TGF β1表达效率最高 ;时效关系显示 ,稳定表达组TGF β1表达活性较瞬时表达组更强。结论 TGF β1基因转染可使BMSCs有效地表达活性TGF β1而产生生物效应。
Objective To investigate the expression of transforming growth factor β1 (TGF β1) gene in bone marrow stromal cells (BMSCs). Methods Six-week-old New Zealand white rabbits BMSCs were subcultured and transfected with 3μl lipofectamine: 1μgpcD NA3 TGFβ1. Immunohistochemical staining and image analysis were used to detect the expression of TGFβ1 Level. The biological activity of TGF-β1 was detected by mink lung epithelial cell growth inhibition assay, and the relationship between dose-effect and time-effect was analyzed. Results The cells in the transient expression group were strongly positive at 48 hours after transfection, and all the cells in the stable expression group were strongly positive for more than 4 weeks. Image analysis showed that the expression of TGFβ1 in transfected cells was significantly increased compared with the control group (P <0.01). The dose-effect relationship showed that the transfection system pcDNA3 TGF β1 (μg): cationic liposome (1) at 1: 3, TGF β1 expression efficiency; aging relationship showed that stable expression group TGF β1 expression was more transiently expressed Group stronger. Conclusion TGF β1 gene transfection can make BMSCs effectively express active TGF β1 and produce biological effects.