The Expression Of Human Insulin Gene Transfer By Chitosan In Vivo And Vitro Of Diabetic Rats

Objective Type 1 diabetes is a kind of auto-immune disease whose key feature is insulin-deficiency and hyperglycemia. There is no other treatment except for injecting ectogenesis insulin. Therefor, transgeneic insulin expression is one of the directions. Our research is to study the expression of human insulin gene transfer by chitosan in vivo and vitro of diabetic rats.Methods An expression vector for human insulin gene was constructed by recombinant gene technique and introduced into fibroblast cells by chitosan-mediated DNA transfection. Following G418 screening, the survived cells were selected and enriched. To assay the transfected cells by immunohistochemical and insulin levels were monitored by radioimmunoassay(RIA). The diabetic model of Wistar rats was duplicated with STZ once intraperitoneally. The recombinant plasmid pCMV. Ins was mixed with chitosan and then injected into the diabetic rats via tail veins. The insulin levels and blood glucose were monitored.Results (1)Extracellular insulin levels in pCMV.Ins transfected group increased significantly as compared with untransfected group(P<0.01) and pCMV transfected group(P<0.01),while there were no changes in extracellular insulin levels between untransfected group and pCMV group(P>0.05). (2)Plasma insulin levels in pCMV.Ins transfected group increased significantly as compared with untransfected group(P<0.01) and pCMV transfected group(P<0.01), while there were no changes in plasma insulin levels between untransfected group and pCMV group(P>0.05).Conclution Human insulin gene was transfected successfully in non-p cells and diabetic rats by chitosan-mediated indicating transgeneic insulin expression would be possible. Chitosan would be a promising vector.