重组热休克蛋白65对NOD小鼠糖尿病的预防作用研究  

Effect of heat shock protein 65 of mycobacterium tuberculosis on prevention of diabetes in nonobese diabetic mice

作　　者：朱爱华[1] 刘景晶[2] 刘文涛[2] 鲁勇[2] 金亮[2] 吴洁[2] 

机构地区：[1]江苏省徐州师范大学生命科学学院,221116 [2]中国药科大学生命科学与技术学院微基因药物实验室

出　　处：《中国实用医药》2007年第36期1-4,共4页China Practical Medicine

基　　金：国家"863"高技术研究发展计划(项目编号:2002AA217031-2);徐州师范大学自然科学研究基金项目(项目编号:05XLA10);博士启动基金项目(项目编号:KY200610)

摘　　要：目的研究重组热休克蛋白65对NOD小鼠糖尿病发生的预防作用,评价其对人1型糖尿病的作用。方法通过PCR方法克隆出结核分枝杆菌的热休克蛋白65的基因序列,构建了pET28a-HSP65高效表达载体,在大肠杆菌中高效可溶性表达。利用硫酸铵分级沉淀、阴离子交换柱层析分离纯化重组蛋白HSP65。在没有任何佐剂存在的情况下使用HSP65免疫非肥胖性糖尿病(NOD)小鼠,通过三次腹腔注射,每月收集被免疫动物的血清,血糖浓度用自动生化分析仪测定。结果HSP65免疫组小鼠血糖平均值及糖尿病的累积发病率和对照组相比均有显著差异(P<0.01)。结论重组蛋白HSP65对NOD小鼠糖尿病的发生有很好的预防作用,可以进一步研究开发这种免佐剂1型糖尿病疫苗。Objective To study the efficacy of heat shock protein 65 kDa(HSP65)of mybobacterium tuberculosis var.bovis in prevention of autoimmune diabetes.Methods The HSP65 gene was derived from Mybobacterium tuberculosis var.bovis genome by PCR and successfully expressed as soluble protein in Escherichia coli.The recombinant protein HSP65 was purified by anion exchange column chromatography and then used to immunize prediabetic NOD mice with three ip inoculations in absence of adjuvants.Serum samples from the immunized mice were collected at monthly interval and the concentration of blood glucose was measured by automatic analyzer.Results Administration of the recombinant protein HSP65 to NOD mice could prevent the development of diabetes efficiently.Conclusion The recombinant protein of HSP65 would be useful for the development of a vaccine against insulin-dependent diabetes mellitus.

关 键 词：热休克蛋白 1型糖尿病 疫苗 血糖 

分 类 号：R587.1[医药卫生—内分泌]