Comparative proteomic analysis to identify the novel target gene of angiotensin II in adrenocortical H295R cells
Comparative proteomic analysis to identify the novel target gene of angiotensin II in adrenocortical H295R cells
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DOI：
10.1507/endocrj.ej20-0144
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发表时间：
2021-01-01
影响因子：
2
通讯作者：
Sugawara, Akira
中科院分区：
文献类型：
--
作者：
Ito, Ryo;Shima, Hiroki;Sugawara, Akira
血管紧张素II （angii）是一种众所周知的肽，在高等脊椎动物中维持电解质平衡。肾上腺的angii刺激通过上调醛固酮合成酶（CYP11B2）基因表达，诱导以醛固酮为主的矿化皮质激素的合成。此外，据报道，Ang II激活多种信号通路，如丝裂原活化蛋白激酶（MAPK）和Ca2+信号。尽管Ang II对肾上腺细胞的信号传导有多种影响，但其除醛固酮合成外的生物学意义尚不清楚。在这项研究中，我们尝试使用蛋白质组学方法结合使用细胞培养氨基酸（SILAC）的稳定同位素标记，在人肾上腺H295R细胞中寻找Ang II的新靶基因。有趣的是，我们发现Ang II刺激提高了磷酸果糖激酶型血小板（PFKP）的蛋白和mRNA水平的表达。此外，Ang II对PFKP的反激活依赖于细胞外信号调节激酶（ERK） 1/2的激活。最后，我们观察到Ang II处理促进了H295R细胞的葡萄糖摄取。综上所述，我们在这里发现PFKP是Ang II的一个新的靶基因，这表明Ang II不仅刺激类固醇生成，还影响葡萄糖代谢。
Angiotensin II (Ang II) is a well-known peptide that maintains the balance of electrolytes in the higher vertebrates. Ang II stimulation in the adrenal gland induces the synthesis of mineralocorticoids, mainly aldosterone, through the up regulation of aldosterone synthase (CYP11B2) gene expression. Additionally, it has been reported that Ang II activates multiple signaling pathways such as mitogen-activated protein kinase (MAPK) and Ca2+ signaling. Although Ang II has various effects on the cellular signaling in the adrenal cells, its biological significance, except for the aldosterone synthesis, is still unclear. In this study, we attempted to search the novel target gene(s) of Ang II in the human adrenal H295R cells using a proteomic approach combined with stable isotopic labeling using amino acid in cell culture (SILAC). Interestingly, we found that Ang II stimulation elevated the expression of phosphofructokinase type platelet (PFKP) in both protein and mRNA levels. Moreover, transactivation of PFKP by Ang II was dependent on extracellular-signal-regulated kinase (ERK) 1/2 activation. Finally, we observed that Ang II treatment facilitated glucose uptake in the H295R cells. Taken together, we here identified PFKP as a novel target gene of Ang II, indicating that Ang II not only stimulates steroidogenesis but also affects glucose metabolism.