脂质体介导的rhIGF-1基因对成骨细胞增殖分化的影响

Effect of liposome-mediated rhIGF-1 gene on proliferation and differentiation of osteoblast

目的探讨重组人胰岛素生长因子(rhIGF-1)基因对体外培养大鼠成骨细胞增殖分化的影响。方法采用酶消化法获取大鼠乳鼠颅骨成骨细胞,传至第3代进行成骨细胞鉴定,钙化结节用茜素红(ARS)染色,钙钴法显示成骨细胞中碱性磷酸酶(ALP)的表达。用脂质体法将pcDNA3.1-rhIGF-1质粒转染成骨细胞。倒置显微镜下观察细胞的生长形态,MTT法检测成骨细胞增殖情况,碱性磷酸酶及骨钙素(OCN)测定成骨细胞活性。结果建立了稳定的大鼠成骨细胞模型;pcDNA3.1-rhIGF-1质粒转染组与对照组及空质粒pcDNA3.1组比较成骨细胞增殖旺盛、ALP和OGN的分泌增加,有显著性差异(P<0.01)。结论外源性rhIGF-1基因能够刺激大鼠成骨细胞增殖、分化,促进骨形成。

Objective To study the effects of the insulin-like growth factor-1（IGF-1） gene on proliferation and differentiation of rat ostcoblust in vitro.Methods Ostcoblast were obtained from within 24 hours newborn rats cranium through trypsin and collagenese digestion. Identified the of the third subculture cells ostcoblast quality , the mineralization nodes were stained by Alizarin Red S （ARS）, Gomori method show the expression of alkaline pboaphatase （ALP）in ostcoblust, pcDNA3. 1-rhIGF-1 plasmid were transfected into ostcoblast by lipofectamine. Morphologic changes of ostcoblust were observed, the proliferation were detected by MTT method, the activity of ossification were studied by alkaline pboaphatase （ALP） and osteocatcin（OCN）. Results Established the stable model of rat ostcoblust. Comparing with control and void plasmid group, pcDNA3.1-rhIGF-1 plasmid group obviously promoted the proliferation of ostcoblust and increased ALP and OCN secretion, the differences were significant（ P 〈 0.01） .Conclusion Exogenous rhIGF-1 gene could stimulate proliferation and differentiation of ostcoblust and promote bone formation.