TIGAR mediates the inhibitory role of hypoxia on ROS production and apoptosis in rat nucleus pulposus cells
TIGAR mediates the inhibitory role of hypoxia on ROS production and apoptosis in rat nucleus pulposus cells
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TIGAR介导缺氧对大鼠髓核细胞ROS产生和凋亡的抑制作用
DOI：
10.1016/j.joca.2017.10.007
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发表时间：
2018-01-01
影响因子：
7
通讯作者：
Chen, N.
中科院分区：
文献类型：
--
作者：
Jiang, L-B.;Cao, L.;Chen, N.
目的：低氧可抑制髓核细胞产生活性氧。据报道，TP 53诱导的糖酵解和凋亡调节因子（TIGAR）可抑制氧化应激。我们试图探讨TIGAR在缺氧对ROS产生和细胞凋亡的影响中的作用。方法：采用SD大鼠尾椎穿刺Co 6 -7椎间盘建立椎间盘退变（IDD）模型。采用免疫组化和蛋白质印迹法检测TIGAR在人和SD大鼠髓核组织中的表达。通过蛋白质印迹法分析用缺氧和氯化钴（CoCl 2）处理的大鼠原代NP细胞的TIGAR表达。在用TIGAR siRNA转染TIGAR沉默后，测量凋亡百分比、线粒体和总细胞内ROS水平。使用H2 O2进一步检查TIGAR对氧化应激的影响。结果：TIGAR基因在人和大鼠退化的NP组织中的表达均呈程度依赖性增加，在缺氧条件下TIGAR基因沉默后，NADPH/NADP（+）和GSH/GSSH比值均显著升高。缺氧及缺氧诱导剂CoCl 2可促进大鼠NP细胞TIGAR和P53的表达。TIGAR沉默逆转了缺氧对细胞内和线粒体ROS产生以及凋亡百分比的抑制作用。然而，TIGAR沉默加剧了H2 O2诱导的ROS产生。结论：TIGAR可能通过提高NADPH/NADP（+）和GSH/GSSH比值，介导缺氧对NP细胞ROS产生和细胞凋亡的保护作用。(c)2017由Elsevier Ltd代表国际骨关节炎研究学会发布。
Objective: Hypoxia has been shown to inhibit reactive oxygen species (ROS) production in nucleus pulposus (NP) cells. The TP53-induced glycolysis and apoptosis regulator (TIGAR) has been reported to suppress oxidative stress. We sought to explore the role of TIGAR in the effect of hypoxia on ROS production and apoptosis.Methods: An intervertebral disc degeneration (IDD) model of Sprague-Dawley (SD) rat caudal spine was established by puncturing the Co6-7 disc. TIGAR expression was detected by immunohistochemistry and western blotting in human and SD rat NP tissues of degenerated discs. Rat primary NP cells treated with hypoxia and cobalt chloride (CoCl2) were analyzed by western blotting for TIGAR expression. After TIGAR silence with TIGAR siRNA transfection, apoptosis percentage, mitochondrial and total intracellular ROS levels were measured. H2O2 was used to further check the effects of TIGAR on oxidative stress. Finally, NADPH/NADP(+) and GSH/GSSH ratio were examined after TIGAR silencing under hypoxic conditions and after H2O2 treatment.Results: A degree-dependent increase in TIGAR expression was observed in human and rat degenerated NP tissues. Hypoxia and hypoxia-inducer CoCl2 enhanced TIGAR and P53 expressions in rat NP cells. TIGAR silence reversed the inhibitory effects of hypoxia on intracellular and mitochondrial ROS production, as well as apoptosis percentage. However, TIGAR silence aggravated H2O2-induced ROS production. In addition, TIGAR increased NADPH/NADP(+) and GSH/GSSH ratio in NP cells.Conclusions: These results suggested that TIGAR appears to mediate the protective role of hypoxia on ROS production and apoptosis percentage by enhancing NADPH/NADP(+) and GSH/GSSH ratio. (c) 2017 Published by Elsevier Ltd on behalf of Osteoarthritis Research Society International.