Reduced glutathione depletion causes necrosis and sensitization to tumor necrosis factor-α-induced apoptosis in cultured mouse hepatocytes
Reduced glutathione depletion causes necrosis and sensitization to tumor necrosis factor-α-induced apoptosis in cultured mouse hepatocytes
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DOI：
10.1053/jhep.2002.33995
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发表时间：
2002-07-01
期刊：
影响因子：
13.5
通讯作者：
Kaplowitz, N
中科院分区：
文献类型：
--
作者：
Nagai, H;Matsumaru, K;Kaplowitz, N
在培养的小鼠肝细胞中研究了对乙酰氨基酚 (A-PAIP)、马来酸二乙酯 (DEM) 或佛尔酮消耗还原型谷胱甘肽 (GSH) 对细胞死亡模式和对肿瘤坏死因子 (TNF) 诱导的细胞死亡的敏感性的影响。剂量依赖性坏死是单独使用 APAP 时细胞死亡的唯一模式，但添加 TNF-α 会诱导细胞凋亡转向大约一半，而不会改变活力的完全丧失。这种效应在 1 和 5 mmol/L APAP 时可见，但在 10 和 20 mmol/L APAP 时受到抑制。细胞凋亡的转变与 caspase 活性的增加、细胞色素 c 的释放和 DNA 阶梯相关，并且被 caspase 抑制剂抑制。 DEM 和佛尔酮也诱导剂量依赖性坏死。在这些条件下用 TNF-α 处理，在 0.25 和 0.5 mmol/L DEM 以及 0.1 和 0.2 mmol/L 佛尔酮下，会导致细胞凋亡形式的增量细胞死亡。在 1.0 和 2.0 mmol/L DEM 和 0.5 mmol/L 佛尔酮时，观察到 90% 至 100% 的坏死，且对 TNF-α 效应具有抵抗力。通过 DNA 梯带和 caspase 抑制剂的抑制证实了 TNF-a 加 DEM 的细胞凋亡。然而，在存在半胱天冬酶抑制剂的情况下，TNF-α诱导的细胞死亡的增加持续表现为坏死的增加。抗氧化剂、维生素 E 和丁基羟基甲苯 (BHT) 的组合可显着抑制单独的 APAP 或 DEM 诱导的坏死，但对 TNF-α 诱导的细胞凋亡的敏感性不受影响。还原型谷胱甘肽单乙酯 (GSH-EE) 可防止坏死和细胞凋亡。总之，A.PA.P、DEM 或佛尔酮消耗 GSH 会导致氧化应激诱导的坏死，并使氧化应激独立的 TNF-α 诱导的细胞凋亡变得敏感。
The effect of reduced glutathione (GSH) depletion by acetaminophen (A-PAIP), diethylmaleate (DEM), or phorone on the mode of cell death and susceptibility to tumor necrosis factor (TNF)-induced cell death was studied in cultured mouse hepatocy-tes. Dose-dependent necrosis was the exclusive mode of cell death with APAP alone, but the addition of TNF-alpha induced a switch to about half apoptosis without changing total loss of viability. This effect was seen at 1 and 5 mmol/L but was inhibited at 10 and 20 mmol/L APAP. The switch to apoptosis was associated with increased caspase activities, release of cytochrome c, and DNA laddering and was inhibited by caspase inhibitors. DEM and phorone also induced dose-dependent necrosis. Treatment with TNF-a under these conditions lead to incremental cell death in the form of apoptosis at 0.25 and 0.5 mmol/L DEM and 0.1 and 0.2 mmol/L phorone. At 1.0 and 2.0 mmol/L DEM and 0.5 mmol/L phorone, 90% to 100% necrosis was observed with resistance to TNF-a effects. The apoptosis with TNF-a plus DEM was confirmed by DNA laddering and inhibition by caspase inhibitors. However, in the presence of caspase inhibitors, the increment in cell death induced by TNF-alpha persisted as an increase in necrosis. A combination of antioxidants, vitamin E, and butylated hydroxytoluene (BHT) markedly inhibited necrosis induced by APAP or DEM alone, but the sensitization to TNF-alpha-induced apoptosis was unaffected. GSH monoethylester (GSH-EE) protected against necrosis and apoptosis. In conclusion, depletion of GSH by A.PA.P, DEM, or phorone causes oxidative stress-induced necrosis and sensitizes to an oxidative stress independent TNF-alpha-induced apoptosis.