Acrolein causes inhibitor κB-independent decreases in nuclear factor κB activation in human lung adenocarcinoma (A549) cells
Acrolein causes inhibitor κB-independent decreases in nuclear factor κB activation in human lung adenocarcinoma (A549) cells
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DOI：
10.1074/jbc.274.14.9200
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发表时间：
1999-04-02
影响因子：
4.8
通讯作者：
Kehrer, JP
中科院分区：
文献类型：
--
作者：
Horton, ND;Biswal, SS;Kehrer, JP
丙烯醛是一种高度亲电子的α，β-不饱和醛，人类在各种情况下都会接触到它。本研究观察了亚致死剂量丙烯醛对人肺腺癌细胞A549核因子-κ B（NF-κ B）活化的影响。在暴露于45 fmol丙烯醛/细胞30分钟后，谷胱甘肽（GSH）和DNA合成以及NF-κ B结合仅减少80%以上。治疗后8小时，所有参数均恢复至正常或超正常水平。用丙烯醛预处理完全阻断了12-O-十四酰基佛波醇-13-乙酸酯（TPA）诱导的NF-κ B活化。用1 mM马来酸二乙酯（DEM）处理1 h的细胞显示GSH和DNA合成分别减少34%和53%，DEM还在处理后2 h将NF-κ B活化减少64%，在8 h恢复到对照的22%以内。丙烯醛和DEM在用TPA处理后2 h均使NF-κ B功能减少50%，如分泌型碱性磷酸酶报告基因测定所示。谷胱甘肽：在DEM处理后8小时恢复到对照水平，但增殖保持显著抑制24小时。有趣的是，DEM引起NF-κ B结合的显著降低，即使在对GSH几乎没有影响的低至0.125 mM的剂量下。丙烯醛和DEM对磷酸化或非磷酸化的抑制剂κ B-α（I κ B-α）的水平都没有任何影响。此外，丙烯醛降低了在环己酰亚胺存在下TPA刺激消耗I κ B-α的细胞中的NF-κ B活化，表明NF-κ B活化的降低不是抑制性蛋白结合增加的结果。这一结论进一步得到以下发现的支持：在用去污剂从I κ B化学解离之前，丙烯醛修饰了胞质溶胶中的NF-κ B。总之，这些数据支持丙烯醛和DEM对NF-κ B活化的抑制是I κ B非依赖性的结论。该机制似乎与NF-κ B亚基中巯基的直接修饰有关。
Acrolein is a highly electrophilic alpha,beta-unsaturated aldehyde to which humans are exposed in various situations. In the present study, the effects of sublethal doses of acrolein on nuclear factor kappa B (NF-kappa B) activation in A549 human lung adenocarcinoma cells were investigated. Immediately following a 30-min exposure to 45 fmol of acrolein/cell, glutathione (GSH) and DNA synthesis and NF-kappa B binding mere reduced by more than 80%. All parameters returned to normal or supranormal levels by 8 h post-treatment. Pretreatment with acrolein completely blocked 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced activation of NF-kappa B. Cells treated for 1 h with 1 mM diethyl maleate (DEM) showed a 34 and 53% decrease in GSH and DNA synthesis, respectively, DEM also reduced NF-kappa B activation by 64% at 2 h post-treatment, with recovery to within 22% of control at 8 h, Both acrolein and DEM decreased NF-kappa B function similar to 50% at 2 h after treatment with TPA, as shown by a secreted alkaline phosphatase reporter assay. GSH: returned to control levels by 8 h after DEM treatment, but proliferation remained significantly depressed for 24 h, Interestingly, DEM caused a profound decrease in NF-kappa B binding, even at doses as low as 0.125 mM that had little effect on GSH. Neither acrolein nor DEM had any effect on the levels of phosphorylated or nonphosphorylated inhibitor kappa B-alpha: (I kappa B-alpha), Furthermore, acrolein decreased NF-kappa B activation in cells depleted of I kappa B-alpha by TPA stimulation in the presence of cycloheximide:, demonstrating that the decrease in NF-kappa B activation was not the result of increased binding by the inhibitory protein. This conclusion was further supported by the finding that acrolein modified NF-kappa B in the cytosol prior to chemical dissociation from I kappa B with detergent, Together, these data support the conclusion that the inhibition of NF-kappa B activation by acrolein and DEM is I kappa B-independent. The mechanism appears to be related to direct modification of thiol groups in the NF-kappa B subunits.