Phosphatidylinositol 3 kinase pathway and 4-hydroxy-2-nonenal-induced oxidative injury in the RPE.
Phosphatidylinositol 3 kinase pathway and 4-hydroxy-2-nonenal-induced oxidative injury in the RPE.
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DOI：
10.1167/iovs.08-2439
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发表时间：
2009-02
影响因子：
4.4
通讯作者：
Cai J
中科院分区：
文献类型：
--
作者：
Chen J;Wang L;Chen Y;Sternberg P;Cai J
4-羟基-2-壬烯醛（4-HNE）是视网膜和视网膜色素上皮中的主要脂质过氧化产物。本研究的目的是探讨NF-E2相关因子2（Nrf 2）和磷脂酰肌醇3（PI 3 K）通路如何影响培养的人视网膜色素上皮（RPE）细胞对4-HNE的反应。用不同浓度的4-HNE和PI 3 K抑制剂LY 294002处理培养的ARPE-19细胞。细胞内谷胱甘肽（GSH）的测定采用高效液相色谱法（HPLC）。双荧光素酶法检测报告质粒瞬时转染后Nrf 2的转录活性。通过实时RT-PCR定量谷氨酸半胱氨酸连接酶（GCL）的mRNA水平。通过免疫沉淀和Western印迹分析来测量HNE加合物在热休克同源蛋白70（Hsc 70）上的形成。用4-HNE处理培养的RPE细胞以剂量依赖性方式增加Nrf 2活性和GSH合成。GCL的调节亚基被4-HNE上调。LY 294002预处理后，抗氧化反应在很大程度上被消除。当抑制PI 3 K时，4-HNE对Hsc 70的修饰作用增强。Nrf 2依赖性抗氧化反应保护免受4-HNE毒性，并且这种保护机制依赖于PI 3 K途径的功能。
4-Hydroxy-2-nonenal (4-HNE) is a major lipid peroxidation product in the retina and the retinal pigment epithelium. The purpose of the present study was to investigate how NF-E2–related factor-2 (Nrf2) and phosphatidylinositol 3 (PI3K) pathways affect the responses of cultured human retinal pigment epithelial (RPE) cells to 4-HNE. Cultured ARPE-19 cells were treated with different concentrations of 4-HNE and a PI3K inhibitor, LY294002. Intracellular glutathione (GSH) was measured by high-performance liquid chromatography (HPLC). The transcriptional activity of Nrf2 was measured by dual luciferase assay after transient transfection with reporter plasmids. The mRNA level of glutamate cysteine ligase (GCL) was quantified by real-time RT-PCR. Formation of HNE adduct on heat shock cognate protein 70 (Hsc70) was measured by immunoprecipitation and Western blot analyses. Treatment with 4-HNE increased Nrf2 activity and GSH synthesis in a dose-dependent manner in cultured RPE cells. The modulatory subunit of GCL was upregulated by 4-HNE. Antioxidant responses were largely abolished by pretreatment with LY294002. The modification of Hsc70 by 4-HNE was increased when PI3K was inhibited. The Nrf2-dependent antioxidant response protects against 4-HNE toxicity, and this protective mechanism is dependent on the functions of the PI3K pathway.
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影响因子：
4.4
作者：
Ethen, Cheryl M.;Reilly, Cavan;Ferrington, Deborah A.
通讯作者：
Ferrington, Deborah A.
影响因子：
4.8
作者：
He, Xiaoqing;Chen, Michael G.;Ma, Qiang
通讯作者：
Ma, Qiang
影响因子：
3.8
作者：
Knoll, N;Ruhe, C;Pool-Zobel, BL
通讯作者：
Pool-Zobel, BL
DOI：
10.1073/pnas.172398899
发表时间：
2002-09-03
影响因子：
11.1
作者：
Dinkova-Kostova, AT;Holtzclaw, WD;Talalay, P
通讯作者：
Talalay, P
影响因子：
5.3
作者：
Cullinan, SB;Gordan, JD;Diehl, JA
通讯作者：
Diehl, JA