دورية أكاديمية
Delivery of Soluble Heme Oxygenase 1 Cell-Penetrating Peptide into Liver Cells in in vitro and ex vivo Models of Cold Ischemia.
| العنوان: | Delivery of Soluble Heme Oxygenase 1 Cell-Penetrating Peptide into Liver Cells in in vitro and ex vivo Models of Cold Ischemia. |
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| المؤلفون: | Venkatachalam AB; Atlantic Centre for Transplantation Research, Dalhousie University, Halifax, NS, Canada., Livingstone SM, Hu Q, Ray A, Wood C, Cimen S, Alwayn IP |
| المصدر: | European surgical research. Europaische chirurgische Forschung. Recherches chirurgicales europeennes [Eur Surg Res] 2017; Vol. 58 (1-2), pp. 51-68. Date of Electronic Publication: 2016 Nov 12. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Karger Country of Publication: Switzerland NLM ID: 0174752 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1421-9921 (Electronic) Linking ISSN: 0014312X NLM ISO Abbreviation: Eur Surg Res Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Basel, New York, Karger. |
| مواضيع طبية MeSH: | Cell-Penetrating Peptides* , Cytoprotection*, Heme Oxygenase-1/*administration & dosage , Liver/*cytology , Reperfusion Injury/*prevention & control, Amino Acid Sequence ; Animals ; Base Sequence ; Cold Ischemia ; Drug Delivery Systems ; Drug Evaluation, Preclinical ; Hep G2 Cells ; Humans ; In Vitro Techniques ; Liver Transplantation ; Molecular Sequence Data ; Perfusion ; Rats, Wistar ; Recombinant Proteins |
| مستخلص: | Background/purpose: Liver transplantation is the treatment of choice in patients with end-stage liver disease. During liver transplantation, ischemia-reperfusion injury (IRI) occurs, which is an inevitable consequence of the transplantation process. To reduce the extent of cellular injury, one of the proteins that have been extensively investigated is heme oxygenase 1 (HO-1), which plays an important role in protecting the organs against IRI. The aim of this study was to introduce an active and functional HO-1 protein conjugated to a cell-penetrating peptide (CPP) in vitro and ex vivo into liver cells in hypothermic and anoxic conditions and to assert its cytoprotective effects. Methods: We generated an enzymatically active soluble (s)HO-1-CPP recombinant protein. The ability of the sHO-1-CPP protein to penetrate McA-RH7777, Clone 9, and Hep G2 cells, primary hepatocytes, and Kupffer and human umbilical vein endothelial cells in vitro, as well as its ability to penetrate a whole liver ex vivo under hypothermic and anoxic conditions, was assessed. An in vitro hypoxia-reoxygenation (HR) model was used to determine the cytoprotective effect of the sHO-1-CPP protein. Results: We showed that our recombinant protein sHO-1-CPP can cross cell membranes into rodent and human liver cells in vitro, and the results were further validated ex vivo, where rodent livers were perfused with an organ preservation solution supplemented with sHO-1-CPP under anoxic and hypothermic conditions. Immunohistochemistry revealed an intracellular localization of sHO-1-CPP in zones 1-3 of the perfused livers. The CPP did not exert any significant toxicity on the cells. Treating cells with sHO-1-CPP showed significant cytoprotection in the in vitro HR model. Conclusions: Our findings show that the recombinant protein sHO-1-CPP can be successfully delivered to cells of a whole organ in an ex vivo hypothermic and anoxic perfusion model and that it provides cytoprotection to hepatocytes in an in vitro HR model. These results hold great potential for future repair and protection of donor organs. Future experiments are planned to confirm these data in in vivo models of IRI. (© 2016 S. Karger AG, Basel.) |
| المشرفين على المادة: | 0 (Cell-Penetrating Peptides) 0 (Recombinant Proteins) EC 1.14.14.18 (Heme Oxygenase-1) |
| تواريخ الأحداث: | Date Created: 20161114 Date Completed: 20170705 Latest Revision: 20170705 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1159/000451079 |
| PMID: | 27838689 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1421-9921 |
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| DOI: | 10.1159/000451079 |