Protection of rat islet viability following heme oxygenase-1 gene transfection via adenoviral vector in vitro1

ObjectiveTo investigate the effect of Heme oxygenase-1 (HO-1) gene transfection on the viability of cultured rat islets, and to explore the potential value of HO-1 gene in islet transplantation.MethodsRecombinant adenovirus vector containing human HO-1 gene (Ad-HO-1) or enhanced green fluorescent protein gene (Ad-EGFP) was generated by using AdEasy system respectively. The rat islets were transfected with Ad-HO-1, Ad-EGFP or blank vector and then cultured for 7 days. Transfection was confirmed by expression of EGFP and human HO-1 protein detected by fluorescence photographs and western blot, respectively. The insulin release upon different concentration of glucose stimulation was detected using insulin radioimmunoassay kit, and stimulation index (SI) was calculated. Glucose-stimulated insulin release was used to assess islet viability.ResultsAdenovirus vector successfully transferred HO-1 gene to rat islet cells in vitro, and the insulin release upon high level of glucose stimulation and stimulation index (SI) of Ad-HO-1-infected islets were significantly higher than those of Ad-EGFP-infected islets and control islets (P < 0.05).ConclusionAdenovinis-mediated HO-1 gene transfection is a feasible strategy to confer cytoprotection and therefore protect the viability of cultured rat islets.