新型有序纳米介孔生物活性玻璃对成骨细胞IGF-Ⅱ基因调控的影响

The influence of mesoporous bioactive glass on the IGF-Ⅱ gene expression of osteoblasts

目的研究新型有序纳米介孔生物活性玻璃的体外生物活性及其对鼠成骨细胞的IGF-Ⅱ基因表达效果,探讨其可能的调控机制。方法定量选取新型的生物活性玻璃MBG(80S15C),浸泡于MEM培养液中,取离子浸出液培养大鼠成骨细胞,使用MTT法和PNPP法测定成骨细胞增殖和ALP的活性,RT-PCR法测定IGF-Ⅱ的mRNA表达,ELISA法测定IGF-Ⅱ蛋白和IGFBP的浓度。结果实验组成骨细胞的ALP活性为对照组的126%,IGF-Ⅱ基因表达为对照组的126%,IGF-Ⅱ蛋白和IGFBP的浓度分别是对照组175%和237%,细胞增殖为对照组的92%。结论MBG的离子浸出液能够明显促进成骨细胞的分化,其机制可能是通过诱导激活IGF-Ⅱ基因的表达,增加IGF-Ⅱ蛋白,IGFBP的合成和分泌,进而促进成骨细胞的分化。

Objective To investigate the bioactivity of mesopo.rous bioactive glass （MBG） and its effect on the IGF-Ⅱ gene expression of rat osteoblast in vitro. Methods Rat osteoblasts were cultured with the ionic products of MBG dissolution in MEM. The cell activity and alkaline phosphatase （ALP） activity of rat osteoblasts were measured with MTT and PNPP. The mRNA expression of IGF-Ⅱ was measured with RT-PCR. The concentration of IGF-Ⅱ protein and IGFBP were measured with ELISA. Results After treatment by the MBG released ions, the ALP activity was increased to 126% of the control concentration of unbound IGF-Ⅱ protein The expression of IGF-Ⅱ was increased to 126%. The and IGF bind protein （IGFBP） was increased to 175% and 237%, respectively. However, the cell activity was only 92% of the control. Conclusions The ionic products of MBG dissolution stimulate the gene expression of IGF-Ⅱ and increase its protein synthesis, which is likely to be responsible for promoting the differentiation of osteoblast obviously, while the cell activity does not change markedly.