细胞穿透肽PEP-1介导增强型绿色荧光蛋白穿透在体小鼠皮肤

目的:研究细胞穿透肽PEP-1介导的增强型绿色荧光蛋白穿透小鼠皮肤的能力。方法:用基因工程的方法制备并纯化EGFP蛋白和PEP-1-EGFP融合蛋白,分别将1mg的蛋白涂抹在昆明小鼠背部皮肤上,4 h后处死小鼠取背部皮肤冰冻切片后立即在倒置荧光显微镜下观察。结果:EGFP处理的小鼠皮肤仅在角质层表面有少量荧光,PEP-1-EGFP融合蛋白处理的小鼠皮肤可见绿色荧光穿透角质层并分布于表皮和真皮内。结论:PEP-1-EGFP融合蛋白能穿透小鼠皮肤角质层并分布于表皮和真皮内,这为将来用细胞穿透肽PEP-1经皮转导大分子药物治疗各种皮肤病以及全身系统性疾病奠定了基础。

Cell-penetrating Peptide PEP-1-mediated Transdermal Delivery of Enhanced Green Fluorescent Protein into Mouse Skin

Objective To investigate the transdermal penetrating ability of the fusion protein PEP-1-EGFP with mouse skin.Methods Two prokaryotic expression plasmids pET15b-EGFP and pET15b-PEP-1-EGFP were constructed and transformed into E.coli BL21(DE3) to express EGFP and fusion protein PEP-1-EGFP,respectively.The expressed EGFP and PEP-1-EGFP were purified with Ni2+-resin affinity chromatography and their capability of transdermal penetrating into mouse skin was visualized using inverted fluorescent microscopy after 4 hour application onto the skin with 1mg of protein.Results The mouse skin applied with EGFP showed little fluorescent on the stratum corneum surface whereas mouse skin applied with PEP-1-EGFP fusion protein showed strong fluorescent both in the epidermis and dermis.Conclusions Cell-penetrating peptide PEP-1-mediated transdermal delivery of enhanced green fluorescent protein into mouse skin provides a basis for the research on topical or transdermal delivery of macromolecular drugs mediated by PEP-1 in the therapy of local or systemic disease.