ABB  Vol.4 No.7 , July 2013
Retrovirus vector transfection of rat insulin gene into pancreas decrease blood glucose of diabetic rat
Author(s) Min-Chuan Lai, Chi Yang*
ABSTRACT

Human and animal diabetes mellitus were controlled by a dietary treatment supplemented with either a sulfonylurea drug or insulin injection. Insulin injections were inconvenient and the hypoglycemia induced by insulin-overdose could be fatal. Sulfonylurea drugs were administered orally, however, do not typically provide 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 human and animal diabetes mellitus. Recently, insulin gene transferred and expressed in non-pancreatic cells as a means for the treatment of diabetes was developed rapidly in the expanding gene therapy. Retrovirus, lentivirus, adenovirus, adenoassociated virus and herpes simplex had been used as viral vectors, and the constructed viral-insulin gene was successfully transferred into diabetic rat cells. A gene, containing promoter, enhancer and rat type I insulin gene (a-chain, b-chain and signal peptide), was constructed into a retrovirus vector in the study. The constructed viral-insulin gene was transferred into mouse fibroblast cell. The insulin concentration in 3-day cultured mouse fibroblast cells was 4806.35 ± 53.72 pg/ml. The insulin concentration for the viral vector containing enhancer and promoter of rat insulin gene was higher than the vector containing only insulin gene by a 61% increase in the cultured mouse fibroblast cells. The enhancer and promoter activity of rat insulin gene would be an important determinant for the expression of insulin gene. The secreted amount of insulin by retrovirus vector contained enhancer/promoter gene in this study could achieve as high concentrations (4806.35 ± 53.72 pg/ml) as the insulin injection therapy. Blood glouse decreased sig-

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


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.
References
[1]   Rabinovitch, A., Suarez-Pinzon, W., Strynadka, K., Ju, Q., Edelstein, D., Brownlee, M., Korbutt, G.S. and Rajotte, R.V. (1999) Transfection of human pancreatic islets with an anti-apoptotic gene (bcl-2) protects beta-cells from cytokine-induced destruction. Diabetes, 48, 12231229. doi:10.2337/diabetes.48.6.1223

[2]   Benoist, C. and Mathis, D. (1997) Cell death mediators in autoimmune diabetes: No shortage of subjects. Cell, 89, 1-3.

[3]   Rand, J.S., Fleeman, L.M., Farrow, H.A., Appleton, D.J. and Lederer, R. (2004) Canine and feline diabetes mellitus: Nature or nurture? Journal of Nutrition, 134, 2072S2080S.

[4]   Leibowitz, G., Beattie, G.M., Kafri, T., Cirulli, V., Lopez, A.D., Hayek A. and Levine, F. (1999) Gene transfer to human pancreatic endocrine cells using viral vectors. Diabetes, 48, 745-753. doi:10.2337/diabetes.48.4.745

[5]   Hazelwood, R.L. and Cieslak, S.R. (1989) In vitro release of pancreatic hormones following 99% pancreatectomy in the chicken. General and Comparative Endocrinology, 73, 308-317. doi:10.1016/0016-6480(89)90104-4

[6]   Maeda, Y., Ikeda, U., Ogasawara, Y., Urabe, M., Takizawa, T., Saito, T., Colosi, P., Kurtzman, G., Shimada, K. and Ozawa, K. (1997) Gene transfer into vascular cells using adeno-associated virus (AAV) vectors. Cardiovascular Research, 35, 514-521.

[7]   Zhang, K., Jin, H., Zhong, F., Li, X., Neng, C., Chen, H., Li, W. and Wen, J. (2012) Adenovirus-mediated canine interferon-gamma expression and its antiviral activity against canine parvovirus. Wei Sheng Wu Xue Bao, 52 (11): 1400-1407. (In Chinese)

[8]   Lau, D., Harte, S.E., Morrow, T.J., Wang, S., Mata, M. and Fink, D.J. (2012) Herpes simplex virus vector-mediated expression of interleukin-10 reduces below-level central neuropathic pain after spinal cord injury. Neurorehabilitation and Neural Repair, 26, 889-897.

[9]   Bogoslovskaia, E.V., Glazkova, D.V., Shipulin, G.A. and Pokrovski?, V.V. (2012) Safety of retroviral vectors in gene therapy. Vestnik Rossiiskoi Akademii Meditsinskikh Nauk, 10, 55-61. (In Russian)

[10]   Kolodka, T.M., Finegold, M., Moss, L. and Woo, S.L. (1995) Gene therapy for diabetes mellitus in rats by hepatic expression of insulin. Proceedings of the National Academy of Sciences of the United States of America, 92, 3293-3297.

[11]   Sugiyama, A., Hattori, S., Tanaka, S., Isoda, F., Kleopoulos, S., Rosenfeld, M., Kaplitt, M., Sekihara, H. and Mobbs, C. (1997) Defective adenoassociated viral-mediated transfection of insulin gene by direct injection into liver parenchyma decrease blood glucose of diabetic mice. Hormone and Metabolic Research, 29, 599-603. doi:10.1055/s-2007-979108

[12]   Ren, B., O’Brien, B.A., Swan, M.A., Koina, M.E., Nassif, N., Wei, M.Q. and Simpson, A.M. (2007) Long-term correction of diabetes in rats after lentiviral hepatic insulin gene therapy. Diabetologia, 50, 1910-1920.

[13]   Tseng, L.N., Tseng, Y.H., Jiang, Y.D., Chang, C.H., Chung, C.H., Lin, B.J., Chuang, L.M., Tai, T.Y. and Sheu, W.H. (2012) Prevalence of hypertension and dyslipidemia and their associations with micro- and macro-vascular diseases in patients with diabetes in Taiwan: An analysis of nationwide data for 2000-2009. Journal of the Formosan Medical Association, 111, 625-636.

[14]   Falqui, L. (1999) Gene therapy for beta-cell functional replacement in IDDM. Journal of Pediatric Endocrinology & Metabolism, 12, 789-793.

 
 
Top