重组蛋白转导结构域-SARA融合蛋白生物学功能的初步研究  

Biological Function of Recombinant Protein Transduction Domain-SARA Fusion Protein

作　　者：加慧卫[1] 黄晨[1] 李晶[2] 张英起[2] 孙世仁[1] 杜锐[1] 

机构地区：[1]第四军医大学西京医院肾脏内科,西安710032 [2]第四军医大学生物技术中心,西安710032

出　　处：《中国生物制品学杂志》2010年第6期581-585,共5页Chinese Journal of Biologicals

摘　　要：目的初步研究重组蛋白转导结构域(Protein transduction domain,PTD)-SARA融合蛋白(PTD-SARA)抗肾脏纤维化的生物学功能。方法采用免疫细胞化学法检测重组PTD-SARA融合蛋白的穿膜作用;通过显微镜观察人肾小管上皮细胞HK2形态学变化,Western blot法测定上皮细胞钙黏蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)和磷酸化Smad3蛋白水平,以比较PTD-SARA融合蛋白和SARA蛋白抑制转化生长因子-β1(TGF-β1)诱发的肾小管上皮细胞向间质细胞转分化作用。结果重组PTD-SARA融合蛋白可高效地转入细胞胞质与胞核;PTD-SARA融合蛋白与SARA蛋白相比,能更显著地上调E-cadherin蛋白的表达,下调α-SMA和磷酸化Smad3蛋白的表达,且差异有统计学意义(P<0.05)。结论与SARA蛋白相比,PTD-SARA融合蛋白可以高效地进入细胞胞质及胞核,并能明显地抑制TGF-β1诱发的肾小管上皮细胞向间质细胞转分化作用,为进一步研究重组PTD-SARA融合蛋白防治肾脏纤维化奠定了基础。Objective To preliminarily study the biological function of recombinant protein transduction domain（PTD）SARA fusion protein in inhibiting renal fibrosis.Methods The transmembrane efficiency of recombinant PTD-SARA fusion protein was determined by immunocytochemical assay.The morphology of HK2 cells was observed by microscopy.The E-cadherin,α-smooth muscle actin（α-SMA）and phospho Smad3 protein levels were determined by Western blot to compare the inhibitory effects of PTDSARA and SARA on TGF-β1-induced renal tubular epithelial to mesenchymal transition.Results Recombinant PTD-SARA fusion protein could enter both cytoplasm and karyon of HK2 cells effectively.Compared with SARA,PTD-SARA up-regulated the expression of E-cadherin and down-regulated those of α-SMA and phosphoSmad3 protein significantly（P〈0.05）.Conclusion Compared with SARA,PTD-SARA may enter the cytoplasm and karyon effectively and inhibit the TGF-β1-induced renal tubular epithelial to mesenchymal transition significantly,which lays a foundation of further study on prevention and treatment of renal fibrosis with recombinant PTD-SARA fusion protein.

关 键 词：SARA 蛋白转导结构域 融合蛋白质类 肾 纤维化 

分 类 号：R733.72[医药卫生—肿瘤]