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Human andanimal diabetes mellitus were controlled by a dietary treatment supplementedwith either a sulfonylurea drug or insulin injection. Insulin injections wereinconvenient and the hypoglycemia induced by insulin-overdose could be fatal.Sulfonylurea drugs were administered orally, however, do not typicallyprovide satisfactory control of blood glucose as a starting treatment in 25% - 30%patients. Therefore, it was imperative to develop a method for the control of humanand animal diabetes mellitus. Recently, insulin gene transferred and expressedin non-pancreatic cells as a means for the treatment of diabetes was developedrapidly in the expanding gene therapy. Retrovirus, lentivirus, adenovirus,adenoassociated virus and herpes simplex had been used as viral vectors, andthe constructed viral-insulin gene was successfully transferred into diabeticrat cells. A gene, containing promoter, enhancer and rat type I insulin genea-chain, b-chain and signal peptide, was constructed into a retrovirus vectorin the study. The constructed viral-insulin gene was transferred into mousefibroblast cell. The insulin concentration in 3-day cultured mouse fibroblast cellswas 4806.35 ± 53.72 pg-ml. The insulin concentration for the viral vectorcontaining enhancer and promoter of rat insulin gene was higher than thevector containing only insulin gene by a 61% increase in the cultured mousefibroblast cells. The enhancer and promoter activity of rat insulin gene wouldbe an important determinant for the expression of insulin gene. The secretedamount of insulin by retrovirus vector contained enhancer-promoter gene inthis study could achieve as high concentrations 4806.35 ± 53.72 pg-ml as theinsulin injection therapy. Blood glouse decreased sig-

nificantly for at last 10 days demonstrated that transfection, direction injection ofviral-insulin gene into pancreas of diabetic rat, was successful. These studiessuggest that the retrovirus vector might be used to transfer the insulin genein vitro and in vivo.



KEYWORDS

Enhancer; Diabetic Rat; Insulin Gene; Promoter; Viral Vector

Cite this paper

Lai, M. and Yang, C. 2013 Retrovirus vector transfection of rat insulin gene into pancreas decrease blood glucose of diabetic rat. Advances in Bioscience and Biotechnology, 4, 769-773. doi: 10.4236-abb.2013.47100.





Autor: Min-Chuan Lai, Chi Yang

Fuente: http://www.scirp.org/



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