摘要
利用基因工程技术高效制备具有治疗2型糖尿病功能的垂体腺苷酸环化酶激活肽(PACAP)衍生物MPL-2,并以2型糖尿病小鼠(db/db小鼠)为模型在体内研究其抗2型糖尿病的生物学作用。实验结果表明:利用基因工程技术制备的PACAP27衍生多肽MPL-2的分子质量为3 902Da.,纯度达97%,其产率可达29.3mg/L发酵产物;在以db/db小鼠为模型的体内葡萄糖耐量实验中,MPL-2可有效促进小鼠胰岛素第一时相(5~15min)分泌,显著提高小鼠的葡萄糖耐受能力。在MPL-2的长效药效学实验中,经过8周连续用药治疗后,MPL-2可显著提高db/db小鼠的胰岛素敏感性,胰岛素耐量实验60min时MPL-2可将小鼠血糖降至初始值的63.52%;同时,在8周连续用药治疗过程中,与生理盐水(NS)处理组相比,MPL-2可有效降低db/db小鼠的体重、空腹血糖、饮食量、饮水量,分别低于NS组21.98%、21.46%、22.20%、60.07%,而且可显著改善db/db小鼠的血脂常数,生物学作用显著优于多肽BAY55-9837。建立了新型基因重组PACAP27衍生多肽MPL-2的高效制备技术,重组多肽MPL-2可有效改善2型糖尿病db/db小鼠的葡萄糖耐量、胰岛素敏感性、血脂常数,显著降低db/db小鼠的体重、空腹血糖、饮食和饮水量,从而发挥治疗2型糖尿病的生物学作用,可为MPL-2的药用研发提供实验数据。
MPL-2, a pituitary adenylate cyclase-activating polypeptide （PACAP） derivative was prepared by gene recombination engineering technique and the effect of anti-type 2 diabetes mellitus was studied in db / db mice. The results showed that the molecular weight of MPL-2 was 3 902Da, and the purity was about 97% with its yeid up to 29.3mg/（ per liter of fermentation product）. The in vivo glucose tolerance test on db / db mice showed that MPL-2 can effectively promote the secretion of insulin in the first phase （5-15min） , and improve glucose tolerance in mice significantly. Furthermore, chronic administration of MPL-2 by daily injection for 8 weeks significantly improves lipid profiles and also greatly increases insulin sensitivity in db / db mice, which decreased the blood glucose of the mice to 63.52% of the initial value at 60min in the insulin tolerance test. At the meanwhile, the body weight, fasting blood glucose, food and water intake of db / db mice were also decreased by MPL-2, which was respectively lower 21.98%, 21.46%, 22.20% and 60.07% than that of NS group. It showed that MPL-2 was more effective than BAY55-9837. In conclusion, the recombinant MPL-2 could effectively improve the glucose tolerance, insulin sensitivity and lipid level of db / db mice, and significantly decreased body weight, fasting blood glucose, food intake and water consumption, thus play a important role in the biological treatment of type 2 diabetes. Experimental basis for the research and development of MPL-2 can also be provided.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2017年第5期59-65,共7页
China Biotechnology
基金
国家自然科学基金项目(81373314)
广东省自然科学基金项目(2015A030313333
2015A030313345)
广东省科技计划项目(2014A020210015
2013B090500105)
中央高校基本科研业务费专项资金(21615412)资助项目
关键词
基因工程
垂体腺苷酸环化酶激活肽衍生物
2型糖尿病
胰岛素敏感性
葡萄糖耐量
Genetic engineering Pituitary adenylatecyclase-activating polypeptide C PACAP） derivativeType 2 diabetes Insulin sensitivity Glucose tolerance
