LIM矿化蛋白1对牙周膜细胞生物学特性的影响

目的以LIM矿化蛋白1（LIM mineralization protein 1,LMP-1）基因构建真核表达载体PET43.1a-LMP-1,转染人牙周膜细胞,观察对其生物学特性的影响。方法采用基因工程技术构建真核表达载体pET43.1a-LMP-1,脂质体法转染人牙周膜细胞,四唑盐比色法和酶联免疫吸附测定法测定真核表达载体pET43.1a-LMP-1转染对牙周膜细胞活性以及碱性磷酸酶（alkaline phosphatase,ALP）的影响。结果与空质粒转染组和空白对照组相比,pET43.1a-LMP-1真核表达载体转染后牙周膜细胞中LMP-1 mRNA和蛋白表达显著增强（P〈0.05）;细胞增殖明显增加;ALP表达明显增强,并随时间延长逐渐上升。结论外源性LMP-1转染进入牙周膜细胞能够促进细胞的增殖和矿化能力。

Effect of Recombinant Plasmid Pet43.1a-LMP-1 on Human Periodontal Ligament Cells

Objective The aim of this study is to construct eukaryotic expression vector PET43.1a-LMP-1 with LIM mineralization protein 1（LMP-1） and investigate the effect of LMP-1 by recombinant plasmids PET43.1a-LMP-1 on human periodontal ligament cells（PDLCs）.Methods The LMP-1 cDNA was cloned into vector PET43.1a for constructing the vector PET43.1a-LMP-1.Periodontal ligament cells were cultured and then transfected with PET43.1a-LMP-1.Expression of LMP-1 mRNA and protein in periodontal ligament cells were detected by RT-PCR and western blot,respectively;cell viabilities were detected by（3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2H-tetrazoliumbromide,MTT） and alkaline phosphatase（ALP） were detected by enzyme linked immunosorbent（ELISA）.Results The expression of LMP-1 mRNA and ALP were detected in PDLCs tranfected with PET43.1a-LMP-1,ALP as well as PDLCs viability were significantly higer than that of the control.Conclusion The results provided a therapeutic strategy for periodontal regeneration with PDLCs transfected PET43.1a-LMP-1.