Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702, South Korea.
Department of Anatomy and Cell Biology, College of Veterinary Medicine, and Research Institute for Veterinary Science, Seoul National University, Seoul 151-742, South Korea.
Free Radic Biol Med. 2014 Apr;69:181-96. doi: 10.1016/j.freeradbiomed.2014.01.006. Epub 2014 Jan 17.
Human carbonyl reductase 1 (CBR1) is a member of the NADPH-dependent short-chain dehydrogenase/reductase superfamily that is known to play an important role in neuronal cell survival via its antioxidant function. Oxidative stress is one of the major causes of degenerative disorders including ischemia. However, the role CBR1 plays with regard to ischemic injury is as yet poorly understood. Protein transduction domains such as PEP-1 are well known and now commonly used to deliver therapeutic proteins into cells. In this study, we prepared PEP-1-CBR1 protein and examined whether it protects against oxidative-stress-induced neuronal cell damage. PEP-1-CBR1 protein was efficiently transduced into hippocampal neuronal HT-22 cells and protected against hydrogen peroxide (H2O2)-induced neuronal cell death. Transduced PEP-1-CBR1 protein drastically inhibited H2O2-induced reactive oxygen species production, the oxidation of intracellular macromolecules, and the activation of mitogen-activated protein kinases, as well as cellular apoptosis. Furthermore, we demonstrated that transduced PEP-1-CBR1 protein markedly protected against neuronal cell death in the CA1 region of the hippocampus resulting from ischemic injury in an animal model. In addition, PEP-1-CBR1 protein drastically reduced activation of glial cells and lipid peroxidation in an animal model. These results indicate that PEP-1-CBR1 protein significantly protects against oxidative-stress-induced neuronal cell death in vitro and in vivo. Therefore, we suggest that PEP-1-CBR1 protein may be a therapeutic agent for the treatment of ischemic injuries as well as oxidative-stress-induced cell damage and death.
人羰基还原酶 1（CBR1）是 NADPH 依赖性短链脱氢酶/还原酶超家族的成员，已知通过其抗氧化功能在神经元细胞存活中发挥重要作用。氧化应激是包括缺血在内的退行性疾病的主要原因之一。然而，CBR1 在缺血性损伤中的作用尚未完全了解。蛋白转导结构域（如 PEP-1）是众所周知的，现在常用于将治疗性蛋白递送到细胞中。在这项研究中，我们制备了 PEP-1-CBR1 蛋白，并研究了它是否对氧化应激诱导的神经元细胞损伤具有保护作用。PEP-1-CBR1 蛋白有效地转导到海马神经元 HT-22 细胞中，并能抵抗过氧化氢（H2O2）诱导的神经元细胞死亡。转导的 PEP-1-CBR1 蛋白可显著抑制 H2O2 诱导的活性氧产生、细胞内大分子的氧化以及丝裂原活化蛋白激酶的激活和细胞凋亡。此外，我们证明，转导的 PEP-1-CBR1 蛋白在动物模型中明显保护了由缺血性损伤引起的海马 CA1 区神经元细胞死亡。此外，PEP-1-CBR1 蛋白在动物模型中还可显著减少小胶质细胞的激活和脂质过氧化。这些结果表明，PEP-1-CBR1 蛋白可显著保护体外和体内氧化应激诱导的神经元细胞死亡。因此，我们认为 PEP-1-CBR1 蛋白可能是治疗缺血性损伤以及氧化应激诱导的细胞损伤和死亡的治疗剂。
