Heat-shock protein B1 upholds the cytoplasm reduced state to inhibit activation of the Hippo pathway in H9c2 cells
Heat-shock protein B1 upholds the cytoplasm reduced state to inhibit activation of the Hippo pathway in H9c2 cells
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热休克蛋白B1维持细胞质还原状态抑制Hippo通路在H9c2细胞中的激活
DOI：
10.1002/jcp.27322
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发表时间：
2019-04-01
影响因子：
5.6
通讯作者：
Zhang, Huali
中科院分区：
文献类型：
--
作者：
Liu, Xiehong;Liu, Ke;Zhang, Huali
热休克蛋白B1（HSPB 1）是一种多功能蛋白质，可保护机体免受氧化应激;然而，其在抗氧化途径中的功能仍不清楚。在这里，我们试图确定HSPB 1在H9 c2细胞受到氧化应激的作用。使用非还原十二烷基硫酸钠聚丙烯酰胺凝胶电泳，我们发现，增加HSPB 1的表达促进还原状态的谷胱甘肽还原酶（GR），过氧化物酶1（Prx 1），硫氧还蛋白1，而敲低HSPB 1衰减这些反应后氧化应激。HSPB 1表达增加促进GR和硫氧还蛋白还原酶的活化。相反，敲低HSPB 1减弱了氧化应激后的这些反应。重要的是，HSPB 1的过表达促进了HSPB 1和氧化Prx 1之间的复合物形成，导致暴露于H2O的H9 c2细胞中的哺乳动物STE 20样激酶1（MST 1）去磷酸化（2），而HSPB 1的下调则诱导了相反的结果。HSPB 1通过增强MST 1的去磷酸化作用调节Hippo通路，导致LATS 1和Yes相关蛋白（雅普）的磷酸化减少。此外，HSPB 1调节YAP依赖的基因表达。因此，HSPB 1促进了H9 c2细胞氧化应激后内源性抗氧化途径的还原状态，并通过调节Hippo信号通路改善了细胞质的氧化还原状态。
Heat-shock protein B1 (HSPB1) is a multifunctional protein that protects against oxidative stress; however, its function in antioxidant pathways remains largely unknown. Here, we sought to determine the roles of HSPB1 in H9c2 cells subjected to oxidative stress. Using nonreducing sodium dodecyl sulfate polyacrylamide gel electrophoresis, we found that increased HSPB1 expression promoted the reduced states of glutathione reductase (GR), peroxiredoxin 1 (Prx1), and thioredoxin 1, whereas knockdown of HSPB1 attenuated these responses following oxidative stress. Increased HSPB1 expression promoted the activation of GR and thioredoxin reductase. Conversely, knockdown of HSPB1 attenuated these responses following oxidative stress. Importantly, overexpression of HSPB1 promoted the complex formation between HSPB1 and oxidized Prx1, leading to dephosphorylation of STE-mammalian STE20-like kinase 1 (MST1) in H9c2 cells exposed to H2O (2), whereas downregulation of HSPB1 induced the opposite results. Mechanistically, HSPB1 regulated the Hippo pathway by enhancing the dephosphorylation of MST1, resulting in reduced phosphorylation of LATS1 and Yes-associated protein (YAP). Moreover, HSPB1 regulated YAP-dependent gene expression. Thus, HSPB1 promoted the reduced state of endogenous antioxidant pathways following oxidative stress in H9c2 cells and improved the redox state of the cytoplasm via modulation of the Hippo signaling pathway.