Cytochrome P450 2E1-derived reactive oxygen species mediate paracrine stimulation of collagen I protein synthesis by hepatic stellate cells
Cytochrome P450 2E1-derived reactive oxygen species mediate paracrine stimulation of collagen I protein synthesis by hepatic stellate cells
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DOI：
10.1074/jbc.m110506200
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发表时间：
2002-03-22
影响因子：
4.8
通讯作者：
Cederbaum, AI
中科院分区：
文献类型：
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作者：
Nieto, N;Friedman, SL;Cederbaum, AI
为了评估依赖CYP2E1的活性氧产生可能的纤维化作用，我们建立了一个模型，使用HepG2细胞（E47细胞）或不（C34细胞）与星状细胞共同培养细胞色素P450 2E1 （CYP2E1）。与单独或与C34细胞共培养相比，与E47细胞共培养的星状细胞细胞内和细胞外H2O2、脂质过氧化和I型胶原蛋白含量均有所增加。胶原蛋白的增加被抗氧化剂和CYP2E1抑制剂所阻止。CYP3A4不模仿CYP2E1的刺激作用。胶原mRNA水平保持不变，脉冲追踪分析表明，两种共培养物的胶原I蛋白半衰期相似。而E47共培养组胶原蛋白合成增加。吡唑处理的大鼠肝细胞（高水平CYP2E1）诱导原代星状细胞中的胶原蛋白，抗氧化剂和CYP2E1抑制剂阻断了这一作用。这些结果表明，cyp2e1衍生的活性氧增加了胶原mRNA的翻译，是E47共培养产生的胶原蛋白增加的原因。这些共培养模型可能有助于理解cyp2e1衍生的ROS对星状细胞功能和激活的影响。
To evaluate possible fibrogenic effects of CYP2E1-dependent generation of reactive oxygen species, a model was developed using co-cultures of HepG2 cells, which do (E47 cells) or do not (C34 cells) express cytochrome P450 2E1 (CYP2E1) with stellate cells. There was an increase in intra- and extracellular H2O2, lipid peroxidation, and collagen type I protein in stellate cells co-cultured with E47 cells compared with stellate cells alone or co-cultured with C34 cells. The increase in collagen was prevented by antioxidants and a CYP2E1 inhibitor. CYP3A4 did not mimic the stimulatory effects found with CYP2E1. Collagen mRNA levels remained unchanged, and pulse-chase analysis indicated similar half-lives of collagen I protein between both co-cultures. However, collagen protein synthesis was increased in E47 co-culture. Hepatocytes from pyrazole-treated rats (with high levels of CYP2E1) induced collagen protein in primary stellate cells, and antioxidants and CYP2E1 inhibitors blocked this effect. These results suggest that increased translation of collagen mRNA by CYP2E1-derived reactive oxygen species is responsible for the increase in collagen protein produced by the E47 co-culture. These co-culture models may be useful for understanding the impact of CYP2E1-derived ROS on stellate cell function and activation.