Oxidative stress increases A1 adenosine receptor expression by activating nuclear factor κB
Oxidative stress increases A1 adenosine receptor expression by activating nuclear factor κB
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DOI：
10.1124/mol.53.4.663
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发表时间：
1998-04-01
影响因子：
3.6
通讯作者：
Ramkumar, V
中科院分区：
文献类型：
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作者：
Nie, ZZ;Mei, Y;Ramkumar, V
A(1) 腺苷受体 (A(1)AR) 在已知会产生活性氧 (ROS) 的条件下有助于腺苷的细胞保护作用。 A(1)AR 表达的药理学操作已被证明可以调节这种细胞保护作用。在这项研究中，我们提供的证据表明，产生的 ROS 可以增加 A(1)AR 的表达，从而抵消 ROS 的有害影响。将 DDT1MF-2 平滑肌细胞与产生 ROS 的化疗剂（例如顺铂 (2.5 μM) 或 H2O2 (10 μM)）一起孵育，在 24 小时内引起 A(1)AR 表达增加。 H2O2 的诱导会被 ROS 清除剂过氧化氢酶减少，但不会被超氧化物歧化酶减少。吡咯烷二硫代氨基甲酸酯 (200 μM)、地塞米松 (100 nM) 或染料木黄酮 (1 μM) 对核因子 kappa B (NF kappa B) 的抑制消除了顺铂介导的 A(1)AR 增加。通过电泳迁移率变动分析和蛋白质印迹检测发现，顺铂促进 NF kappa B（但不是 AP-1）快速易位至细胞核。 A,AR 启动子中推定的 NF kappa B 序列有效地与标记的 KB 探针竞争结合源自 DDT1MF-2 细胞的核制剂。用与萤火虫荧光素酶报告基因偶联的 A,AR 启动子瞬时转染 DDT1MF-2 细胞，产生顺铂诱导和吡咯烷二硫代氨基甲酸盐敏感的荧光素酶活性，表明 A(1)AR 启动子序列中存在功能性 NF kappa B 结合位点。用（R）-苯基异丙基腺苷（1μM）（A（1）AR的激动剂）处理细胞，减少了顺铂介导的脂质过氧化，在阻断A（1）AR后，这种情况被逆转。这些数据表明，ROS 可以通过激活 A(1)AR 基因上的 NF kappa B 调节位点来增加该基因的表达，从而增强腺苷的细胞保护作用。
The A(1) adenosine receptor (A(1)AR) contributes to the cytoprotective action of adenosine under conditions known to generate reactive oxygen species (ROS). Pharmacological manipulation of A(1)AR expression has been shown to modulate this cytoprotective role. In this study, we provide evidence that ROS generated could increase the expression of the A(1)AR and thereby offset the detrimental effects of ROS. Incubation of DDT1MF-2 smooth muscle cells with ROS-generating chemotherapeutic agents, such as cisplatin (2.5 mu M) or H2O2 (10 mu M), elicited an increase in A(1)AR expression within 24 hr. The induction by H2O2 was reduced by the ROS scavenger catalase but not superoxide dismutase. Inhibition of nuclear factor kappa B (NF kappa B) by pyrrolidine dithiocarbamate (200 mu M), dexamethasone (100 nM), or genistein (1 mu M) abrogated the cisplatin-mediated increase in A(1)AR. Cisplatin promoted rapid translocation of NF kappa B (but not AP-1) to the nucleus, as detected by electrophoretic mobility shift assays and by Western blotting. A putative NF kappa B sequence in the A,AR promoter effectively competed with labeled KB probe for binding in nuclear preparations derived from DDT1MF-2 cells. Transient transfection of DDT1MF-2 cells with the A,AR promoter coupled to firefly luciferase reporter gene led to cisplatin-inducible and pyrrolidine dithiocarbamate-sensitive luciferase activity, suggesting the presence of functional NF kappa B binding site(s) in the A(1)AR promoter sequence. Treatment of cells with (R)-phenylisopropyladenosine (1 mu M), an agonist of the A(1)AR, reduced cisplatin-mediated lipid peroxidation, which was reversed after blockade of the A(1)AR. These data suggest that ROS can increase the expression of the A(1)AR by activating NF kappa B regulatory site(s) on this gene and thereby enhance the cytoprotective role of adenosine.