Lee Jae Hyuk, Kim Kyuseok, Jo You Hwan, Hwang Ji Eun, Chung Hea Jin, Yang Chungmi
Department of Emergency Medicine, Seoul National University Bundang Hospital, Gyeonggi-do, Republic of Korea.
Department of Emergency Medicine, Seoul National University Bundang Hospital, Gyeonggi-do, Republic of Korea.
J Surg Res. 2018 Jul;227:88-94. doi: 10.1016/j.jss.2017.11.005. Epub 2018 Mar 14.
Ischemia/reperfusion injury is characterized by excess generation of reactive oxygen species (ROS). The purpose of this study is to test the effect of reoxygenation speed on ROS production and the cellular injury responses in hypoxic macrophages RAW 264.7 cells and its potential mechanisms for the generation of ROS.
After hypoxic exposure of RAW 264.7 cells for 20 h, reoxygenation was performed for 6 h by stepwise increase in oxygen concentration (0.8% increase of oxygen every 15 min) in the slow reoxygenation (SRox) group or by moving the culture flasks quickly to a normoxic incubator in the rapid reoxygenation (RRox) group. To identify the potential effect of reoxygenation speed on the generation of ROS, the cells were pretreated with apocynin, VAS2870, and MitoTEMPO before the induction of hypoxia.
SRox significantly decreased cell death and cytotoxicity compared with RRox (P < 0.05). RRox resulted in significantly more generation of ROS, interleukin-1β, interleukin-6, and nitric oxide than SRox (P < 0.05). SRox also increased the expression of prosurvival proteins and decreased apoptosis. In cells pretreated with VAS2870 or MitoTEMPO, the reduced ROS generation by SRox was maintained. However, pretreatment with apocynin abolished the effect of reoxygenation speed on ROS generation.
SRox compared with RRox decreased cellular injury in hypoxic RAW 264.7 cells by decreasing ROS and inflammatory cytokine production and decreasing apoptosis.
缺血/再灌注损伤的特征是活性氧（ROS）过度生成。本研究的目的是测试复氧速度对缺氧巨噬细胞RAW 264.7细胞中ROS产生、细胞损伤反应的影响及其产生ROS的潜在机制。
将RAW 264.7细胞缺氧暴露20小时后，在缓慢复氧（SRox）组中通过逐步增加氧气浓度（每15分钟氧气浓度增加0.8%）进行6小时复氧，或在快速复氧（RRox）组中将培养瓶快速转移至常氧培养箱中进行6小时复氧。为了确定复氧速度对ROS产生的潜在影响，在诱导缺氧前用阿朴吗啡、VAS2870和线粒体靶向抗氧化剂MitoTEMPO对细胞进行预处理。
与RRox相比，SRox显著降低了细胞死亡和细胞毒性（P < 0.05）。RRox导致产生的ROS、白细胞介素-1β、白细胞介素-6和一氧化氮明显多于SRox（P < 0.05）。SRox还增加了促生存蛋白的表达并减少了细胞凋亡。在用VAS2870或MitoTEMPO预处理的细胞中，SRox降低的ROS生成得以维持。然而，用阿朴吗啡预处理消除了复氧速度对ROS生成的影响。
与RRox相比，SRox通过减少ROS和炎性细胞因子的产生以及减少细胞凋亡，降低了缺氧RAW 264.7细胞的细胞损伤。
