大鼠体内生物素示踪重组人胶原蛋白的方法研究

Study on the method of tracing recombinant human collagen by biotin in vivo

目的:建立一种在大鼠体内示踪外部重组人胶原蛋白的方法。方法:采用超级生物素标记重组人胶原蛋白,将标记后的重组人胶原蛋白植入大鼠体内,实验周期结束后取植入部位组织,制备石蜡切片,用辣根过氧化物酶标记链霉亲和素识别病理切片上生物素标记的蛋白,二氨基联苯胺法染色(DAB法染色),显微镜观察组织情况。结果:在病理切片中观察到生物素标记重组人胶原蛋白的棕褐色沉淀,实现了植入组织的重组人胶原蛋白的定位。结论:生物素标记重组人胶原蛋白的方法灵敏度高,操作简单,可快速实现对重组人胶原蛋白的追踪。本文通过设立生物素对照组,排除生物素本身体内代谢对实验结果的影响。

Objective:To establish a method for tracing external recombinant human collagen in rats.Methods:The recombinant human collagen was labeled with super biotin,and the labeled recombinant human collagen was implanted into rats.After the experimental cycle,the tissue of the implanted site was taken and paraffin section was prepared.The biotin labeled protein on pathological section was identified by horseradish peroxidase streptavidin,and the tissue was observed by DAB dyeing.Results:The brown precipitate of the biotin-labeled recombinant human collagen was observed in pathological sections,which realized the localization of recombinant human collagen in implanted tissues.Conclusion:The method of biotin labeling recombinant human collagen has high sensitivity,simple operation,and can quickly realize the tracking of recombinant human collagen.In this paper,the biotin control group has been established to exclude the influence of biotin metabolism on the experimental results.