Involvement of Na/K-ATPase in hydrogen peroxide-induced activation of the Src/ERK pathway in LLC-PK1 cells.
Involvement of Na/K-ATPase in hydrogen peroxide-induced activation of the Src/ERK pathway in LLC-PK1 cells.
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DOI：
10.1016/j.freeradbiomed.2014.03.036
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发表时间：
2014-06
影响因子：
7.4
通讯作者：
Xie Z
中科院分区：
文献类型：
--
作者：
Wang Y;Ye Q;Liu C;Xie JX;Yan Y;Lai F;Duan Q;Li X;Tian J;Xie Z
我们已经证明Na/K-ATPase与Src相互作用。在这里，我们测试的作用，这种相互作用在H2 O2诱导的Src和ERK的激活。我们发现，LLC-PK 1细胞暴露于通过向培养基中加入葡萄糖氧化酶产生的H2 O2激活Src和ERK 1/2。它也引起了适度的减少表面Na/K-ATP酶的数量和哇巴因敏感的Rb+摄取。H_2O_2对LLC-PK_1细胞的上述作用与Na/K-ATP酶的特异性配体哇巴因相似。在α1 Na/K-ATP酶基因敲低的PY-17细胞中，H2 O2对Src和ERK 1/2的作用受到抑制。尽管野生型α1或Src调节缺陷的A420 P突变体α1的表达挽救了PY-17细胞中的泵活性，但只有α1而不是A420 P突变体能够恢复H2 O2诱导的蛋白激酶活化。与此一致，用pNaKtide破坏Na/K-ATP酶/Src复合物的形成减弱了H2 O2对激酶的影响。此外，H2 O2对Na/K-ATP酶介导的Src调节的直接影响被证明。H2 O2通过Na/K-ATP酶/Src介导的信号通路抑制E-cadherin的表达。以上结果提示，Na/K-ATP酶/Src复合物可能是H2 O2调节Src/ERK蛋白激酶进而调节肾上皮细胞表型的受体机制之一。
We have shown that Na/K-ATPase interacts with Src. Here, we test the role of this interaction in H2O2-induced activation of Src and ERK. We found that exposure of LLC-PK1 cells to H2O2 generated by the addition of glucose oxidase into the culture medium activated Src and ERK1/2. It also caused a modest reduction in the number of surface Na/K-ATPases and in ouabain-sensitive Rb+ uptake. These effects of H2O2 seem similar to those induced by ouabain, a specific ligand of Na/K-ATPase, in LLC-PK1 cells. In accordance, we found that the effects of H2O2 on Src and ERK1/2 were inhibited in α1 Na/K-ATPase-knockdown PY-17 cells. Whereas expression of wild-type α1 or the A420P mutant α1 defective in Src regulation rescued the pumping activity in PY-17 cells, only α1, and not the A420P mutant, was able to restore the H2O2-induced activation of protein kinases. Consistent with this, disrupting the formation of the Na/K-ATPase/Src complex with pNaKtide attenuated the effects of H2O2 on the kinases. Moreover, a direct effect of H2O2 on Na/K-ATPase-mediated regulation of Src was demonstrated. Finally, H2O2 reduced the expression of E-cadherin through the Na/K-ATPase/Src-mediated signaling pathway. Taken together, the data suggest that the Na/K-ATPase/Src complex may serve as one of the receptor mechanisms for H2O2 to regulate Src/ERK protein kinases and consequently the phenotype of renal epithelial cells.
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影响因子：
4.8
作者：
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