Anti-inflammatory effect of transduced PEP-1-heme oxygenase-1 in Raw 264.7 cells and a mouse edema model
| Autor: | Hoon Jae Jeong, Suman Dutta, Dae Won Kim, Eun Jeong Sohn, Duk-Soo Kim, Young Nam Kim, Hyun Sook Hwang, Soon Won Kwon, Jinseu Park, Eun Hee Ahn, Mi Jin Kim, Won Sik Eum, Soo Young Choi |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Male
Recombinant Fusion Proteins Cell Biophysics Inflammation Biochemistry Cell Line Mice chemistry.chemical_compound Western blot Transduction Genetic medicine Animals Edema Molecular Biology Heme RAW 264.7 Cells Mice Inbred ICR medicine.diagnostic_test Anti-Inflammatory Agents Non-Steroidal Lymphokine Cell Biology Fusion protein Molecular biology Protein Structure Tertiary Heme oxygenase Disease Models Animal medicine.anatomical_structure chemistry medicine.symptom Heme Oxygenase-1 |
| Zdroj: | Biochemical and Biophysical Research Communications. 411:354-359 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2011.06.147 |
| Popis: | Highlights: {yields} Recombinant PEP-1 heme oxygenase-1 expression vector was constructed and overexpressed. {yields} We investigated transduction efficiency of PEP-1-HO-1 protein in Raw 264.7 cells. {yields} PEP-1-HO-1 was efficiently transduced into Raw 264.7 cells in a dose and time dependent manner. {yields} PEP-1-HO-1 exerted anti-inflammatory activity in Raw 264.7 cells and in a mice edema model. {yields} PEP-1-HO-1 could be used as a therapeutic drug against inflammatory diseases. -- Abstract: Heme oxygenase-1 (HO-1), which catalyzes the degradation of free heme to biliverdin, carbon monoxide (CO), and free iron (Fe{sup 2+}), is up-regulated by several cellular stress and cell injuries, including inflammation, ischemia and hypoxia. In this study, we examined whether fusion of HO-1 with PEP-1, a protein transduction domain that is able to deliver exogenous molecules to living cells or tissues, would facilitate HO-1 delivery to target cells and tissues, and thereby effectively exert a therapeutically useful response against inflammation. Western blot analysis demonstrated that PEP-1-HO-1 fusion proteins were transduced into Raw 264.7 cells in time- and dose-dependent manners, and were stably maintained in the cells for about 60 h. In addition, fluorescence analysis revealed that only PEP-1-HO-1 fusion proteins were significantly transduced into the cytoplasm of cells, while HO-1 proteinsmore » failed to be transduced. In lipopolysaccharide (LPS)-stimulated Raw 264.7 cells and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse edema model, transduced PEP-1-HO-1 fusion proteins effectively inhibited the overexpression of pro-inflammatory mediators and cytokines. Also, histological analysis demonstrated that PEP-1-HO-1 remarkably suppressed ear edema. The results suggest that the PEP-1-HO-1 fusion protein can be used as a therapeutic molecule against reactive oxygen species-related inflammatory diseases.« less |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect