肽-DNA纳米颗粒用于HIF-1αΔODD转染脊髓损伤动物模型的研究

Investigation on the effectiveness of peptide-DNA nanoparticles mediating HIF-1αΔODDtransfection to SCI animal model

为了探讨肽-DNA纳米颗粒技术用于转染脊髓损伤动物模型的可能性,本研究在克隆得到氧浓度非敏感性的HIF-1α突变体基因(HIF-1αΔODD)并构建其真核重组表达载体(pEGFPC1-HIF-1αΔODD)的基础上,构建用于转染动物模型的肽-DNA纳米颗粒,将其转染脊髓损伤大鼠模型,用组织免疫荧光方法检测内/外源性HIF-1α在动物模型中的表达情况.结果显示,外源性HIF-1α在实验组脊髓组织中正确表达,说明构建的pEGFPC1-HIF-1αΔODD的肽-DNA纳米颗粒成功转染了稳定的脊髓损伤大鼠模型,肽-DNA纳米颗粒技术可作为中枢神经系统损伤动物模型的分子干预研究的一种新的选择.

To investigate the effectiveness of peptide-DNA nanoparticles mediating HIF-1αΔODD transfecting to SCI animal model,peptide-DNA nanoparticles were prepared to transfect pEGFPC1-HIF-1αΔODD into SCI（spinal cord injury） model of rats on the basis of cloning the oxygen-insensitive gene HIF-1αΔODD and then successfully constructing the recombinant eukaryotic expression vector pEGFPC1-HIF-1αΔODD in this study.Then immunofluorescence technique was used to detect the expression of endogenous and exogenous HIF-1α in SCI animal model.Results showed that exogenous HIF-1α protein was expressed only in the intervention group and total HIF-1α protein was expressed more in the intervention group than in the control group,which showed that the peptide-DNA nanoparticles of HIF-1αΔODD transfected SCI model of rats successfully.The technique of peptide-DNA nanoparticles can be employed to transfect the target gene into the animal model and even may be an alternative candidate for animal model intervention study at molecule level in vivo.