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Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
Sponsored by:Sichuan Society of Zoologists; Chengdu Giant Panda Breeding Research Foundation; Sichuan Association of Wildlife Conservation; Sichuan University
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Your Position :Home->Past Journals Catalog->2019 Vol.38 No.6

Continuous Expression of Single-Strand Insulin Analogue in Skeletal Muscles for the Long-Term Efficient Gene Therapy of Type 1 Diabetic Mice
Author of the article:YANG Ping, SU Ruoke, DENG Lu, YANG Yueyao, WANG Gang*
Author's Workplace:Engineering Research Center in Biomaterials, Sichuan University, Chengdu 610064, China
Key Words:insulin analogue; Pluronic L64-electropulse system; type 1 diabetic mice; skeletal muscle gene therapy
Abstract:Objective To investigate the treatment effects of naked plasmid containing single-strand insulin gene via Pluronic L64-electropulse system (L/E) on type 1 diabetes. Methods C-peptide sequences of original insulin gene was replaced by linker (GGGGS)3, and pcDNA-INS plasmid was constructed based on pcDNA3.1(+). BALB/c diabetic mice were established by streptozotocin (STZ) via intraperitoneal injection, and divided into 3 groups:Group pcDNA-INS was treated by pcDNA-INS naked plasmid, Group L/E-pcDNA3.1(+) was treated by L/E system combined with pcDNA3.1(+) naked plasmid treatments, and Group L/E-pcDNA-INS was treated by L/E system combined with pcDNA-INS naked plasmid treatments. All these groups were injected with corresponding plasmids and treated by L/E system after 1 hour according to therapy protocols, and the mice of control group (Group NC) were injected with the same volume of saline in tibialis anterior muscles on both sides. Blood glucose, serum insulin and other indices were detected. Results The pcDNA-INS plasmid was constructed and identified. The STZ-diabetic mice models with higher blood glucose and lower body mass than those of Group NC mice were also successfully established (P<0.05). Group L/E-pcDNA-INS showed lower blood glucose level and higher serum insulin content than both Group pcDNA-INS and Group L/E-pcDNA3.1(+)(P<0.05), and all the other indices such as survival rates, HE staining and immunohistochemical results, and appearances were much closer to the Group NC mice. Moreover, the immunohistochemical results at the end of therapy reflected that the pancreas insulin content of all diabetic mice was still much lower than normal mice, while the muscle insulin content of the L/E-pcDNA-INS-treated mice was much adequate, and the survival rates of L/E-pcDNA-INS-treated mice were 100%(12/12)(vs. Group NC, P > 0.05). Conclusion Insulin analogue gene combined with Pluronic L64-electropulse system apparently improve the remission of type 1 diabetic mice, and thus provide some experimental cues for the future related gene therapy.
2019,38(6): 607-615 收稿日期:2019-05-17
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