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Construction of a Fusion Peptide 5rolGLP-HV and Analysis of its Therapeutic Effect on Type 2 Diabetes Mellitus and Thrombosis in Mice

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Abstract

Glucagon-like peptide-1 (GLP-1), is currently used to treat type 2 diabetes mellitus and hirudin (HV), plays an important role in controlling thrombosis and cardiovascular diseases. This investigation aimed to develop a fusion peptide 5rolGLP-HV which combined functions of rolGLP-1 and rHV to treat diabetes and thrombosis. In this study, we constructed a fusion gene including five copies of rolGLP-1 and one copy of rHV (5rolGLP-HV). The optimum expression conditions of 5rolGLP-HV in a soluble form were 0.8 mM IPTG induction when OD600 reached 0.6–0.8 and further growing at 25 °C for 9 h. Isolated rolGLP-1 and rHV were acquired by trypsin digestion in vitro, and the concentration of them was determined by HPLC in vivo. Oral administration of 5rolGLP-HV significantly decreased the levels of blood glucose, GHbA1C, TC, and TG in diabetic mice at the time of 3 weeks compared to the saline-treated group (p < 0.05), while the insulin level was reversed significantly (p < 0.05). 5rolGLP-HV treatment significantly shortened the length of thrombus in thrombosis mice compared to the saline-treated group (p < 0.01). These results indicated that 5rolGLP-HV had dual-function in treating diabetes and preventing thrombosis.

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Abbreviations

T2DM:

Type 2 diabetes mellitus

DPP-IV:

Dipeptidyl peptidase IV

rolGLP-1:

Recombinant oral long-acting GLP-1

HV:

Hirudin

STZ:

Streptozotocin

AUC:

Area under the curve

OD:

Optical density

OGTT:

Oral glucose tolerance test

GHbA1C:

Glycosylated hemoglobin A1c

TC:

Total cholesterol

TG:

Triglyceride

FBG:

Fasting blood-glucose

ATU:

Antithrombin units

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Acknowledgments

This study is supported by the Key Technologies R&D Program of Tianjin (14ZCZDSY00013).

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Author notes

  1. Minggang Li

    Present address: Life Science College, Nankai University, No. 94, Weijin Road, Nankai District, 300071, Tianjin, China

Authors and Affiliations

  1. State Key Laboratory of Medicinal Chemical Biology, Nankai University, 300071, Tianjin, China

    Zaizhong Ni, Yaofang Zhang, Haisong Wang, Yiming Wei, Baicheng Ma, Junfeng Hao, Peipei Tu, Huikun Duan, Xiaodan Li, Pingzhe Jiang, Xiaofeng Ma, Bin Wang, Ri Wu & Minggang Li

  2. Key Laboratory for Bioactive Materials of the Ministry of Education, Institute of Molecular Biology, College of Life Science, Nankai University, 300071, Tianjin, China

    Zaizhong Ni, Yaofang Zhang, Haisong Wang, Yiming Wei, Baicheng Ma, Junfeng Hao, Peipei Tu, Huikun Duan, Xiaodan Li, Pingzhe Jiang, Xiaofeng Ma, Bin Wang, Ri Wu & Minggang Li

  3. Department of Preventive Medicine, Wenzhou Medical University, Wenzhou, Zhejiang, 325035, China

    Jianhong Zhu

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Ni, Z., Zhang, Y., Wang, H. et al. Construction of a Fusion Peptide 5rolGLP-HV and Analysis of its Therapeutic Effect on Type 2 Diabetes Mellitus and Thrombosis in Mice. Appl Biochem Biotechnol 179, 59–74 (2016). https://doi.org/10.1007/s12010-016-1979-x

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