Intracellular glutathione levels are involved in carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone-induced apoptosis in As4.1 juxtaglomerular cells
Intracellular glutathione levels are involved in carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone-induced apoptosis in As4.1 juxtaglomerular cells
复制标题
DOI：
10.3892/ijmm.2011.604
复制
发表时间：
2011-04-01
影响因子：
5.4
通讯作者：
Park, Woo Hyun
中科院分区：
文献类型：
--
作者：
Han, Yong Hwan;Park, Woo Hyun
羰基氰对三氟甲氧基苯腙（FCCP）是真核细胞线粒体氧化磷酸化的解偶联剂.在本研究中，我们研究了活性氧（ROS）和谷胱甘肽（GSH）在FCCP诱导的As4.1肾小球细胞死亡的参与。FCCP在早期（10-150 min）降低细胞内ROS水平，48 h升高。FCCP通过下调Mn-SOD蛋白表达而抑制Mn-SOD活性。Ebselen（一种抗氧化剂）显著减弱FCCP处理的细胞中的ROS水平，但不能阻止FCCP诱导的细胞死亡。此外，细胞内GSH含量在FCCP处理的10分钟内迅速减少，这伴随着线粒体膜电位[MMP（Δ φ m）]的降低。GSH合成抑制剂L-丁硫基乙硫醚亚砜亚胺（BSO）能显著增加FCCP对As4.1细胞的致死作用。然而，N-乙酰半胱氨酸（NAC，GSH前体和抗氧化剂）减弱GSH耗竭，MMP（64，m）的损失和FCCP处理的As4.1细胞的细胞死亡。此外，NAC增加Mn-SOD活性，降低FCCP处理的As4.1细胞中的ROS水平。总之，这些结果表明，ROS水平相比，细胞内GSH水平与FCCP诱导的As4.1肾小球细胞凋亡更密切相关。
Carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone (FCCP) is an uncoupler of mitochondria] oxidative phosphorylation in eukaryotic cells. In the present study, we investigated the involvement of reactive oxygen species (ROS) and glutathione (GSH) in FCCP-induced As4.1 juxtaglomerular cell death. Intracellular ROS levels were decreased by FCCP at the early time points (10-150 min) and increased at 48 h. FCCP inhibited the activity of Mn-superoxide dismutase (Mn-SOD) via down-regulating its protein expression. Ebselen (an antioxidant) significantly attenuated ROS levels in FCCP-treated cells, but did not prevent FCCP-induced cell death. Moreover, intracellular GSH content was rapidly diminished within 10 min of FCCP treatment, which was accompanied by a reduction of the mitochondrial membrane potential [MMP (Delta phi m)]. L-buthionine sulfoximine (BSO, a GSH synthesis inhibitor) significantly augmented As4.1 cell death by FCCP. However, N-acetylcysteine (NAC, a GSH precursor and antioxidant) attenuated GSH depletion, MMP (64,m) loss and cell death in FCCP-treated As4.1 cells. In addition, NAC increased Mn-SOD activity and decreased ROS levels in FCCP-treated As4.1 cells. In conclusion, these results suggest that compared to ROS levels, intracellular GSH levels are more closely linked to FCCP-induced apoptosis in As4.1 juxtaglomerular cells.