LIM矿化蛋白1对牙周膜细胞生物学特性的影响  

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

作　　者：俞少杰[1] 付云[1] 赵川江[1] 李颖[1] 邓雨泉[1] 

机构地区：[1]中山大学附属口腔医院牙周科,广东广州510055

出　　处：《广东牙病防治》2011年第11期585-589,共5页Journal of Dental Prevention and Treatment

基　　金：广东省医学科学技术研究基金(A2009225);广东省科技计划项目(2010B031100010)

摘　　要：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。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.

关 键 词：LIM矿化蛋白1 基因转染 人牙周膜细胞 

分 类 号：R781.4[医药卫生—口腔医学]