Nucleotide Sequencing, Cloning, and Expression of Capra hircus Heme Oxygenase-1 in Caprine Islets to Promote Insulin Secretion In Vitro
| Autor: | Faezeh Vakhshiteh, Zahra Ajdari, Zeenathul Nazariah Allaudin, Mohd Azmi Mohd Lila, Sahar Abbasiliasi |
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| Rok vydání: | 2014 |
| Předmět: |
Male
endocrine system DNA Complementary endocrine system diseases medicine.medical_treatment Green Fluorescent Proteins Molecular Sequence Data Bioengineering Biology Transfection Applied Microbiology and Biotechnology Biochemistry Islets of Langerhans Rapid amplification of cDNA ends Complementary DNA Insulin Secretion medicine Animals Insulin Cloning Molecular Molecular Biology geography geography.geographical_feature_category Goats Pancreatic islets Sequence Analysis DNA Flow Cytometry Islet Molecular biology Heme oxygenase Transplantation medicine.anatomical_structure Pancreas Heme Oxygenase-1 Biotechnology |
| Zdroj: | Molecular Biotechnology. 57:75-83 |
| ISSN: | 1559-0305 1073-6085 |
| Popis: | Transplantation of islets of Langerhans that have been isolated from whole pancreas is an attractive alternative for the reversal of Type 1 diabetes. However, in vitro culture of isolated pancreatic islets has been reported to cause a decrease in glucose response over time. Hence, the improvement in islet culture conditions is an important goal in islet transplantation. Heme Oxygenase-1 (HO-1) is a stress protein that has been described as an inducible protein with the capacity of preventing apoptosis and cytoprotection via radical scavenging. Therefore, this study was aimed to assess the influence of endogenous HO-1 gene transfer on insulin secretion of caprine islets. The full-length cDNA sequence of Capra hircus HO-1 was determined using specific designed primers and rapid amplification of cDNA ends of pancreatic tissue. The HO-1 cDNA was then cloned into the prokaryotic expression vectors and transfected into caprine islets using lipid carriers. Efficiency of lipid carriers to transfect caprine islets was determined by flow cytometry. Insulin secretion assay was carried out by ovine insulin ELISA. The finding demonstrated that endogenous HO-1 gene transfer could improve caprine islet function in in vitro culture. Consequently, strategies using HO-1 gene transfer to islets might lead to better outcome in islet transplantation. |
| Databáze: | OpenAIRE |
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