新型基因重组PACAP衍生物MPL-2的制备及其抗2型糖尿病作用研究

doi:10.13523/j.cb.20170508

中国生物工程杂志

2017, Vol. 37

Issue (5): 59-65 DOI: 10.13523/j.cb.20170508

技术与方法

新型基因重组PACAP衍生物MPL-2的制备及其抗2型糖尿病作用研究

王得华¹, 马义¹, 韩磊¹, 肖兴¹, 李艳伟¹, 党诗莹¹, 范志勇², 文涛¹, 洪岸¹

1. 暨南大学细胞生物学系暨南大学生物医药研究院广东省生物工程药物重点实验室, 基因工程药物国家工程研究中心广州 510632;
2. 广州中医药大学第二附属医院(广东省中医院) 广州 510405

Preparation of Novel Recombinant PACAP Derivative MPL-2 and Its Effect on Anti-type 2 Diabetes Mellitus

WANG De-hua¹, MA Yi¹, HAN Lei¹, XIAO Xing¹, LI Yan-wei¹, DANG Shi-ying¹, FAN Zhi-yong², WEN Tao¹, HONG An¹

1. Department of Cell Biology of Jinan University, Institute of Biological Medicine of Jinan University, Guangdong Provincial Key Laboratory of Bioengineering Medicine, National Engineering Research Center of Genetic Medicine, Guangzhou 510632, China;
2. The Second Affiliated Hospital of Guangzhou University of Chinese Medicine(Guangdong Provincial Hospital of Traditional Chinese Medicine), Guangzhou 510405, China

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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。

关键词： 垂体腺苷酸环化酶激活肽衍生物; 胰岛素敏感性; 2型糖尿病; 基因工程; 葡萄糖耐量

Abstract:

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.

Key words: Type 2 diabetes Insulin sensitivity Glucose tolerance Genetic engineering Pituitary adenylatecyclase-activating polypeptide(PACAP) derivative

收稿日期: 2017-01-03 出版日期: 2017-05-25

ZTFLH:

Q78

基金资助:

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

通讯作者: 马义 E-mail: tmayi@jnu.edu.cn

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引用本文:

王得华, 马义, 韩磊, 肖兴, 李艳伟, 党诗莹, 范志勇, 文涛, 洪岸. 新型基因重组PACAP衍生物MPL-2的制备及其抗2型糖尿病作用研究[J]. 中国生物工程杂志, 2017, 37(5): 59-65.

WANG De-hua, MA Yi, HAN Lei, XIAO Xing, LI Yan-wei, DANG Shi-ying, FAN Zhi-yong, WEN Tao, HONG An. Preparation of Novel Recombinant PACAP Derivative MPL-2 and Its Effect on Anti-type 2 Diabetes Mellitus. China Biotechnology, 2017, 37(5): 59-65.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170508 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I5/59

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