Transduction of PEP-1-heme oxygenase-1 into insulin-producing INS-1 cells protects them against cytokine-induced cell death.
Pro-inflammatory cytokines play a crucial role in the destruction of pancreatic β-cells, thereby triggering the development of autoimmune diabetes mellitus. We recently developed a cell-permeable fusion protein, PEP-1-heme oxygenase-1 (PEP-1-HO-1) and investigated the anti-inflammatory effects in macrophage cells. In this study, we transduced PEP-1-HO-1 into INS-1 insulinoma cells and examined its protective effect against cytokine-induced cell death. PEP-1-HO-1 was successfully delivered into INS-1 cells in time- and dose-dependent manner and was maintained within the cells for at least 48 h. Pre-treatment with PEP-1-HO-1 increased the survival of INS-1 cells exposed to cytokine mixture (IL-1β, IFN-γ, and TNF-α) in a dose-dependent manner. PEP-1-HO-1 markedly decreased cytokine-induced production of reactive oxygen species (ROS), nitric oxide (NO), and malondialdehyde (MDA). These protective effects of PEP-1-HO-1 against cytokines were correlated with the changes in the levels of signaling mediators of inflammation (iNOS and COX-2) and cell apoptosis/survival (Bcl-2, Bax, caspase-3, PARP, JNK, and Akt). These results showed that the transduced PEP-1-HO-1 efficiently prevented cytokine-induced cell death of INS-1 cells by alleviating oxidative/nitrosative stresses and inflammation. Further, these results suggested that PEP-1-mediated HO-1 transduction may be a potential therapeutic strategy to prevent β-cell destruction in patients with autoimmune diabetes mellitus.
Department of Physiology, College of Medicine, Hallym University, Chunchon 200-702, Republic of Korea.
Su Jin
SJ
Lee
Department of Physiology, College of Medicine, Hallym University, Chunchon 200-702, Republic of Korea.
Hyung Kyung
HK
Kang
Department of Pharmacology, College of Medicine, Hallym University, Chunchon 200-702, Republic of Korea.
Dong Keun
DK
Song
Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Republic of Korea.
Won Sik
WS
Eum
Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Republic of Korea.
Jinseu
J
Park
Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Republic of Korea. Electronic address: sychoi@hallym.ac.kr.
Soo Young
SY
Choi
Department of Physiology, College of Medicine, Hallym University, Chunchon 200-702, Republic of Korea. Electronic address: hykwon@hallym.ac.kr.
Hyeok Yil
HY
Kwon
D000818:Animals; D016923:Cell Death / Q000502:physiology*; D045744:Cell Line, Tumor; D016207:Cytokines / Q000502:physiology*; D051547:Heme Oxygenase-1 / Q000235:genetics*; D007328:Insulin / Q000096:biosynthesis*; D007340:Insulinoma / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D008315:Malondialdehyde / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D014161:Transduction, Genetic
D016207:Cytokines; D007328:Insulin; D009573:Nitrites; D017382:Reactive Oxygen Species; D011993:Recombinant Fusion Proteins; D008315:Malondialdehyde; D051547:Heme Oxygenase-1; C561500:PEP-1-heme oxygenase-1 fusion protein
Cytokines; PEP-1-HO-1; Transduction; Type 1 diabetes; β-Cell destruction