长效促胰岛素降糖酵母的构建及其对糖尿病模型小鼠的治疗效果  

Construction of yeast strains expressing long-acting glucagon-like peptide-1(GLP-1) and their therapeutic effects on type 2 diabetes mellitus mouse model

作　　者：吴日[1] 马超[1] 李晓丹[1] 段会坤 姬艳丽[1] 王宇[1] 姜苹哲 王海松[1] 屠培培[1] 李淼[1] 尼钢钢[1] 马百成[2] 李明刚[1] 

机构地区：[1]南开大学生命科学学院,教育部生物活性材料重点实验室,天津300071 [2]天津市儿童医院,天津300074

出　　处：《遗传》2015年第2期183-191,共9页Hereditas（Beijing)

基　　金：天津市科技支撑重点计划项目(编号:14ZCZDSY00013)资助

摘　　要：益生菌生物药物是指通过口服表达药用多肽(蛋白)的重组益生菌活细胞达到治疗疾病的新型口服给药系统。为了构建一种能有效防治2型糖尿病的酵母生物药物,文章首先构建了酿酒酵母(S.cerevisiae)整合型表达载体p NK1-PGK,并且通过绿色荧光蛋白(GFP)证明其表达功能正常,利用该载体将10?GLP-1(Glucagon-like peptide-1)基因转化到酿酒酵母INVSc1中,通过营养缺陷型和Western blotting成功筛选出表达10?GLP-1的长效促胰岛素降糖酵母(Long-acting GLP-1 hypoglycemic yeast,LHY)。该酵母生长迅速,外源基因10?GLP-1表达稳定,表达量达到1.56 mg/g细胞湿重。通过链脲佐菌素和高脂高糖饮食联合诱导的方法构建了2型糖尿病小鼠模型,用LHY对其进行口服灌胃治疗,证明LHY具有较好疗效,明显降低血糖水平。Probiotics, i.e., bacteria expressing therapeutic peptides （protein）, are used as a new type of orally ad- ministrated biologic drugs to treat diseases. To develop yeast strains which could effectively prevent and treat type 2 diabetes mellitus, we firstly constructed the yeast integrating plasmid pNK1-PGK which could successfully expressgreen fluorescent protein （GFP） in Saccharomyces cerevisiae. The gene encoding ten tandem repeats ot glucagon-nKe peptide-1（10 × GLP-1） was cloned into the vector pNK1-PGK and the resulting plasmids were then transformed into the S. cerevisiae INVScl. The long-acting GLP-1 hypoglycemic yeast （LHY） which grows rapidly and expresses 10×GLP-1 stably was selected by nutrition screening and Western blotting. The amount of 10×GLP-1 produced by LHY reached 1.56 mg per gram of wet cells. Moreover, the oral administration of LHY significantly reduced blood glucose level in type 2 diabetic mice induced by streptozotocin plus high fat and high sugar diet.

关 键 词：2型糖尿病 胰高血糖素样肽-1(GLP-1) 长效促胰岛素降糖酵母(LHY) 重组酿酒酵母 糖尿病小鼠模型 

分 类 号：R587.1[医药卫生—内分泌] R-332[医药卫生—内科学]