Kim Mi Jin, Jeong Hoon Jae, Kim Dae Won, Sohn Eun Jeong, Jo Hyo Sang, Kim Duk-Soo, Kim Hyun Ah, Park Eun Young, Park Jong Hoon, Son Ora, Han Kyu Hyung, Park Jinseu, Eum Won Sik, Choi Soo Young
Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon, Gangwondo, Korea.
Department of Biochemistry and Molecular Biology, Research Institute of Oral Sciences, College of Dentistry, Kangnung-Wonju National University, Gangneung, Gangwondo, Korea.
PLoS One. 2014 Jan 23;9(1):e86034. doi: 10.1371/journal.pone.0086034. eCollection 2014.
Paraoxonase 1 (PON1) is an antioxidant enzyme which plays a central role in various diseases. However, the mechanism and function of PON1 protein in inflammation are poorly understood. Since PON1 protein alone cannot be delivered into cells, we generated a cell permeable PEP-1-PON1 protein using protein transduction domains, and examined whether it can protect against cell death in lipopolysaccharide (LPS) or hydrogen peroxide (H2O2)-treated Raw 264.7 cells as well as mice with 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced skin inflammation. We demonstrated that PEP-1-PON1 protein transduced into Raw 264.7 cells and markedly protected against LPS or H2O2-induced cell death by inhibiting cellular reactive oxygen species (ROS) levels, the inflammatory mediator's expression, activation of mitogen-activated protein kinases (MAPKs) and cellular apoptosis. Furthermore, topically applied PEP-1-PON1 protein ameliorates TPA-treated mice skin inflammation via a reduction of inflammatory response. Our results indicate that PEP-1-PON1 protein plays a key role in inflammation and oxidative stress in vitro and in vivo. Therefore, we suggest that PEP-1-PON1 protein may provide a potential protein therapy against oxidative stress and inflammation.
对氧磷酶1（PON1）是一种抗氧化酶，在多种疾病中发挥核心作用。然而，人们对PON1蛋白在炎症中的机制和功能了解甚少。由于PON1蛋白本身无法进入细胞，我们利用蛋白质转导结构域生成了一种可穿透细胞的PEP-1-PON1蛋白，并研究它是否能保护脂多糖（LPS）或过氧化氢（H2O2）处理的Raw 264.7细胞以及12-O-十四酰佛波醇-13-乙酸酯（TPA）诱导的皮肤炎症小鼠免受细胞死亡。我们证明，PEP-1-PON1蛋白可转导进入Raw 264.7细胞，并通过抑制细胞活性氧（ROS）水平、炎症介质的表达、丝裂原活化蛋白激酶（MAPKs）的激活和细胞凋亡，显著保护细胞免受LPS或H2O2诱导的细胞死亡。此外，局部应用PEP-1-PON1蛋白可通过减轻炎症反应改善TPA处理的小鼠皮肤炎症。我们的结果表明，PEP-1-PON1蛋白在体外和体内的炎症和氧化应激中起关键作用。因此，我们认为PEP-1-PON1蛋白可能为抗氧化应激和炎症提供一种潜在的蛋白质疗法。
