Ahn Eun Hee, Kim Dae Won, Shin Min Jea, Ryu Eun Ji, Yong Ji In, Chung Seok Young, Cha Hyun Ju, Kim Sang Jin, Choi Yeon Joo, Kim Duk-Soo, Cho Sung-Woo, Lee Keunwook, Cho Yoon Shin, Kwon Hyeok Yil, Park Jinseu, Eum Won Sik, Choi Soo Young
Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chuncheon, Gangwon‑do 24252, Republic of Korea.
Department of Biochemistry and Molecular Biology, Research Institute of Oral Sciences, College of Dentistry, Gangneung-Wonju National University, Gangneung, Gangwon-do 25457, Republic of Korea.
Int J Mol Med. 2016 Jul;38(1):217-24. doi: 10.3892/ijmm.2016.2599. Epub 2016 May 20.
Antioxidant 1 (ATOX1) functions as an antioxidant against hydrogen peroxide and superoxide, and therefore may play a significant role in many human diseases, including diabetes mellitus (DM). In the present study, we examined the protective effects of Tat-ATOX1 protein on streptozotocin (STZ)-exposed pancreatic insulinoma cells (RINm5F) and in a mouse model of STZ-induced diabetes using western blot analysis, immunofluorescence staining and MTT assay, as well as histological and biochemical analysis. Purified Tat-ATOX1 protein was efficiently transduced into RINm5F cells in a dose- and time-dependent manner. Additionally, Tat-ATOX1 protein markedly inhibited reactive oxygen species (ROS) production, DNA damage and the activation of Akt and mitogen activated protein kinases (MAPKs) in STZ-exposed RINm5F cells. In addition, Tat-ATOX1 protein transduced into mice pancreatic tissues and significantly decreased blood glucose and hemoglobin A1c (HbA1c) levels as well as the body weight changes in a model of STZ-induced diabetes. These results indicate that transduced Tat-ATOX1 protein protects pancreatic β-cells by inhibiting STZ-induced cellular toxicity in vitro and in vivo. Based on these findings, we suggest that Tat-ATOX1 protein has potential applications as a therapeutic agent for oxidative stress-induced diseases including DM.
抗氧化蛋白1（ATOX1）具有对抗过氧化氢和超氧化物的抗氧化作用，因此可能在包括糖尿病（DM）在内的多种人类疾病中发挥重要作用。在本研究中，我们使用蛋白质免疫印迹分析、免疫荧光染色和MTT法，以及组织学和生化分析，研究了Tat-ATOX1蛋白对链脲佐菌素（STZ）处理的胰腺胰岛素瘤细胞（RINm5F）的保护作用，以及在STZ诱导的糖尿病小鼠模型中的作用。纯化的Tat-ATOX1蛋白以剂量和时间依赖性方式有效地转导到RINm5F细胞中。此外，Tat-ATOX1蛋白显著抑制STZ处理的RINm5F细胞中活性氧（ROS）的产生、DNA损伤以及Akt和丝裂原活化蛋白激酶（MAPKs）的激活。此外，转导到小鼠胰腺组织中的Tat-ATOX1蛋白显著降低了STZ诱导的糖尿病模型中的血糖和糖化血红蛋白（HbA1c）水平以及体重变化。这些结果表明，转导的Tat-ATOX1蛋白通过在体外和体内抑制STZ诱导的细胞毒性来保护胰腺β细胞。基于这些发现，我们认为Tat-ATOX1蛋白作为包括DM在内的氧化应激诱导疾病的治疗剂具有潜在应用价值。
