title: Anti-inflammatory effect of transduced PEP-1-heme oxygenase-1 in Raw 264.7 cells and a mouse edema model.
abstract: Heme oxygenase-1 (HO-1), which catalyzes the degradation of free heme to biliverdin, carbon monoxide (CO), and free iron (Fe(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 60h. In addition, fluorescence analysis revealed that only PEP-1-HO-1 fusion proteins were significantly transduced into the cytoplasm of cells, while HO-1 proteins 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.
authors: Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Republic of Korea.
authors: Soon Won
authors: SW
authors: Kwon
authors: Eun Jeong
authors: EJ
authors: Sohn
authors: Dae Won
authors: DW
authors: Kim
authors: Hoon Jae
authors: HJ
authors: Jeong
authors: Mi Jin
authors: MJ
authors: Kim
authors: Eun Hee
authors: EH
authors: Ahn
authors: Young Nam
authors: YN
authors: Kim
authors: Suman
authors: S
authors: Dutta
authors: Duk-Soo
authors: DS
authors: Kim
authors: Jinseu
authors: J
authors: Park
authors: Won Sik
authors: WS
authors: Eum
authors: Hyun Sook
authors: HS
authors: Hwang
authors: Soo Young
authors: SY
authors: Choi
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000627:therapeutic use*; D002460:Cell Line; D004195:Disease Models, Animal; D004487:Edema / Q000188:drug therapy / Q000628:therapy*; D051547:Heme Oxygenase-1 / Q000235:genetics / Q000627:therapeutic use*; D007249:Inflammation / Q000188:drug therapy / Q000628:therapy*; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000627:therapeutic use*; D014161:Transduction, Genetic
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D011993:Recombinant Fusion Proteins; D051547:Heme Oxygenase-1; C561500:PEP-1-heme oxygenase-1 fusion protein