Adenosine A1 receptor-mediated transactivation of the EGF receptor produces a neuroprotective effect on cortical neurons in vitro
Adenosine A1 receptor-mediated transactivation of the EGF receptor produces a neuroprotective effect on cortical neurons in vitro
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DOI：
10.1038/aps.2009.80
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发表时间：
2009-07
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影响因子：
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通讯作者：
Ke-qiang Xie;Li-min Zhang;Yan-Po Cao;Jun Zhu;Lin-yin Feng
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文献类型：
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作者：
Ke-qiang Xie;Li-min Zhang;Yan-Po Cao;Jun Zhu;Lin-yin Feng
目的：了解腺苷a1受体（a1r）介导表皮生长因子受体（EGFR）转激活的机制。方法：用a1r特异性激动剂N - 6-环戊基腺苷（CPA）处理原代培养的氧糖剥夺大鼠皮质神经元和HEK293/ a1r细胞。Western blot检测磷酸化egfr、Akt、ERK1/2的表达。利用免疫沉淀和免疫细胞化学检测EGFR和a1r之间的相互作用。结果：a1r激动剂CPA可激活蛋白激酶B (Akt)，保护皮层初级神经元免受氧-葡萄糖剥夺（OGD）损伤。a1r和EGFR在神经元膜上共定位并形成免疫复合物。1r刺激通过PI3K和Src激酶信号通路诱导显著的EGFR磷酸化；这种刺激对皮层神经元有神经保护作用。CPA导致皮质神经元细胞外调节激酶1和2 （ERK1/2）的持续磷酸化，但仅在HEK 293/ a1r细胞中发生短暂磷酸化。对a1r激动剂的反应主要通过依赖于百日咳毒素（PTX）敏感的g1蛋白和HEK 293/ a1r中金属蛋白酶的EGFR转激活介导。结论：a1r介导的EGFR转激活在初级皮质神经元中具有神经保护作用。PI3激酶和Src激酶在这一反应中起关键作用。
Aim:To understand the mechanism of the transactivation of the epidermal growth factor receptor (EGFR) mediated by the adenosine A 1 receptor (A 1 R).Methods:Primary cultured rat cortical neurons subjected to oxygen-glucose deprivation (OGD) and HEK293/A 1 R cells were treated with the A 1 R-specific agonist N 6-cyclopentyladenosine (CPA). Phospho-EGFR, Akt, and ERK1/2 were observed by Western blot. An interaction between EGFR and A 1 R was detected using immunoprecipitation and immunocytochemistry.Results:The A 1 R agonist CPA causes protein kinase B (Akt) activation and protects primary cortical neurons from oxygen-glucose deprivation (OGD) insult. A 1 R and EGFR co-localize in the membranes of neurons and form an immunocomplex. A 1 R stimulation induces significant EGFR phosphorylation via a PI3K and Src kinase signaling pathway; this stimulation provides a neuroprotective effect in cortical neurons. CPA leads to sustained phosphorylation of extracellularly regulated kinases 1 and 2 (ERK1/2) in cortical neurons, but only to transient phosphorylation in HEK 293/A 1 R cells. The response to the A 1 R agonist is mediated primarily through EGFR transactivation that is dependent on pertussis toxin (PTX)-sensitive G i protein and metalloproteases in HEK 293/A 1 R.Conclusion:A 1 R-mediated EGFR transactivation confers a neuroprotective effect in primary cortical neurons. PI3 kinase and Src kinase play pivotal roles in this response.