Differential peroxiredoxin hyperoxidation regulates MAP kinase signaling in human articular chondrocytes
Differential peroxiredoxin hyperoxidation regulates MAP kinase signaling in human articular chondrocytes
复制标题
DOI：
10.1016/j.freeradbiomed.2019.01.005
复制
发表时间：
2019-04-01
影响因子：
7.4
通讯作者：
Loeser, Richard F.
中科院分区：
文献类型：
--
作者：
Collins, John A.;Wood, Scott T.;Loeser, Richard F.
过氧化物氧还蛋白（Peroxiredoxin，Prx）家族的半胱氨酸依赖性过氧化物酶控制细胞内过氧化氢的水平，并可以调节信号转导。通过过氧化等机制抑制Prx，导致氧化应激条件，可改变稳态信号传导。为了确定Prx 1-Prx 3氧化对人软骨细胞中MAP激酶和IGF-1信号传导事件的影响，本研究使用2-甲基-1，4-萘醌（甲萘醌）和2，3-二甲基-1，4-萘醌（DMNQ）作为H2 O2生成工具，因为它们的作用机制不同。甲萘醌和DMNQ产生类似水平的细胞内H2 O2测定使用的生物传感器Orp 1-roGFP和通过测量Prx氧化还原状态。然而，甲萘醌产生更高水平的线粒体H2 O2与Prx 3过氧化和磷酸化的Prx 1，而DMNQ治疗与细胞溶质Prx 1和Prx 2，但不是线粒体Prx 3的过氧化。甲萘醌和DMNQ都诱导p38持续磷酸化，但只有DMNQ激活JNK。甲萘醌而不是DMNQ抑制IGF-1诱导的Akt磷酸化。用腺病毒载体转导以过表达Prx 3的软骨细胞显示出对甲萘醌的反应减少的PrxSO（2/3）形成，这与IGF-1介导的Akt信号传导的恢复和p38磷酸化的抑制有关。Prx 1和Prx 2过表达对Prx氧化还原状态没有影响，但Prx 1过表达增强了基础Akt磷酸化。这些结果表明，特定的Prx亚型的过氧化与不同的细胞信号转导事件，并确定Prx 3氧化还原状态作为一个重要的调节剂的合成代谢和分解代谢信号转导。有针对性的策略，以防止线粒体Prx 3过氧化可能是有用的，在维持细胞的氧化还原平衡和稳态信号。
The peroxiredoxin (Prx) family of Cys-dependent peroxidases control intracellular levels of H2O2 and can regulate signal transduction. Inhibition of the Prxs, through hyperoxidation amongst other mechanisms, leads to oxidative stress conditions that can alter homeostatic signaling. To determine the effects oxidation of Prx1-Prx3 has on MAP kinase and IGF-1 signaling events in human chondrocytes, this study used 2-methyl-1,4-naphthoquinone (menadione) and 2,3-dimethyl-1,4-naphthoquinone (DMNQ) as H2O2-generating tools due to their differential mechanisms of action. Menadione and DMNQ generated similar levels of intracellular H2O2 as determined using the biosensor Orp1-roGFP and by measuring Prx redox status. However, menadione generated higher levels of mitochondrial H2O2 associated with Prx3 hyperoxidation and phosphorylation of Prx1 while DMNQ treatment was associated with hyperoxidation of cytosolic Prx1 and Prx2 but not mitochondrial Prx3. Both menadione and DMNQ induced sustained phosphorylation of p38 but only DMNQ activated JNK. Menadione but not DMNQ inhibited IGF-1-induced Akt phosphorylation. Chondrocytes transduced with an adenoviral vector to overexpress Prx3 displayed decreased PrxSO(2/3) formation in response to menadione which was associated with restoration of IGF-1-mediated Akt signaling and inhibition of p38 phosphorylation. Prx1 and Prx2 overexpression had no effects on Prx redox status but Prx1 overexpression enhanced basal Akt phosphorylation. These results suggest that hyperoxidation of specific Prx isoforms is associated with distinct cell signaling events and identify Prx3 redox status as an important regulator of anabolic and catabolic signal transduction. Targeted strategies to prevent mitochondrial Prx3 hyperoxidation could be useful in maintaining cellular redox balance and homeostatic signaling.