Reactive Oxygen Species Induced by Bile Acid Induce Apoptosis and Protect Against Necrosis in Pancreatic Acinar Cells
Reactive Oxygen Species Induced by Bile Acid Induce Apoptosis and Protect Against Necrosis in Pancreatic Acinar Cells
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DOI：
10.1053/j.gastro.2011.02.054
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发表时间：
2011-06-01
期刊：
影响因子：
29.4
通讯作者：
Criddle, David N.
中科院分区：
文献类型：
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作者：
Booth, David M.;Murphy, John A.;Criddle, David N.
背景与目的：氧化应激与胰腺炎的发病机制有关，但抗氧化剂的临床试验产生了相互矛盾的结果。我们研究了胰腺腺泡细胞损伤中细胞内活性氧的作用。方法：使用共聚焦显微镜研究了新鲜分离的鼠和人胰腺腺泡细胞，以测量细胞内和线粒体ROS浓度（[ROS]（I）和[ROS]（M））、细胞溶质和线粒体钙浓度（[ROS]（I）和[ROS]（M））、细胞内钙浓度（[ROS]（I）和[ROS]（M））、细胞内钙浓度（[ROS]（I）和[ROS]（M））和细胞内钙浓度（[ROS]（M））的变化。（[Ca 2 +]（C）和[Ca 2 +]（M）），烟酰胺腺嘌呤二核苷酸磷酸水平降低，以及响应于牛磺石胆酸酸硫酸盐（TLC-S）或氧化剂甲萘醌的死亡途径。用全细胞膜片钳技术测定钙激活的氯离子电流，有或没有三磷酸腺苷（ATP）。研究结果：TLC-S诱导[Ca 2 +]（C）和[Ca 2 +]（M）的延长增加，导致[ROS]（I）和[ROS]（M）的剂量依赖性增加，ATP的产生受损，细胞凋亡和坏死。2，4-二甲氧基-2-甲基萘抑制抗氧化剂还原的烟酰胺腺嘌呤二核苷酸磷酸奎宁氧化还原酶可增强[ROS]（I）和细胞凋亡的增加，但减少坏死，而抗氧化剂N-乙酰基-L-半胱氨酸可减少[ROS]（I）和细胞凋亡，但增加坏死。抑制线粒体ROS的产生阻止了细胞凋亡，但没有改变坏死;自噬没有检测到的作用。修补的ATP防止[Ca 2 +]（C）和坏死的持续增加。结论：胆汁酸损伤胰腺腺泡细胞时，[ROS]（M）和[ROS]（I）的增加促进细胞凋亡，但不促进细胞坏死。这些结果表明，急性胰腺炎的氧化应激需要抗氧化剂的替代策略。
BACKGROUND & AIMS: Oxidative stress is implicated in the pathogenesis of pancreatitis, but clinical trials of antioxidants have produced conflicting results. We examined the role of intracellular reactive oxygen species (ROS) in pancreatic acinar cell injury. METHODS: Freshly isolated murine and human pancreatic acinar cells were studied using confocal microscopy to measure changes in intracellular and mitochondrial ROS concentrations ([ROS](I) and [ROS](M)), cytosolic and mitochondrial calcium concentrations ([Ca2+](C) and [Ca2+](M)), reduced nicotinamide adenine dinucleotide phosphate levels, and death pathways in response to taurolithocholate acid sulfate (TLC-S) or the oxidant menadione. Ca2+-activated Cl- currents were measured using whole-cell patch clamp, with or without adenosine triphosphate (ATP). RESULTS: TLC-S induced prolonged increases in [Ca2+](C) and [Ca2+](M), which led to dose-dependent increases in [ROS](I) and [ROS](M), impaired production of ATP, apoptosis, and necrosis. Inhibition of the antioxidant reduced nicotinamide adenine dinucleotide phosphate quinine oxidoreductase by 2,4-dimethoxy-2-methylnaphthalene potentiated the increases in [ROS](I) and apoptosis but reduced necrosis, whereas the antioxidant N-acetyl-L-cysteine reduced [ROS](I) and apoptosis but increased necrosis. Inhibition of mitochondrial ROS production prevented apoptosis but did not alter necrosis; autophagy had no detectable role. Patched ATP prevented sustained increases in [Ca2+](C) and necrosis. CONCLUSIONS: Increases in [ROS](M) and [ROS](I) during bile acid injury of pancreatic acinar cells promote apoptosis but not necrosis. These results indicate that alternative strategies to antioxidants are required for oxidative stress in acute pancreatitis.