质膜 Ca2+ ATP 酶在鞘氨醇激酶-1 调节细胞 Ca2+ 平衡中的作用

IF 2.9 4区 医学 Q2 PHYSIOLOGY
Luisa Michelle Volk, Jan-Erik Bruun, Sandra Trautmann, Dominique Thomas, Stephanie Schwalm, Josef Pfeilschifter, Dagmar Meyer Zu Heringdorf
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引用次数: 0

摘要

磷脂酰肌苷-1-磷酸(S1P)是一种无处不在的脂质介质,可通过特定的 G 蛋白偶联受体(GPCR)在细胞内发挥作用。先前的研究表明,在小鼠胚胎成纤维细胞中,S1P 裂解酶的缺失会导致细胞膜[Ca2+]i 的慢性升高,并增强 Ca2+ 的储存。在这里,我们利用两个独立生成的稳定敲除 SphK1(SphK1-KD1/2)的 EA.hy926 细胞系,研究了鞘氨醇激酶(SphK)-1 在 Ca2+ 信号转导中的作用。在 SphK1-KD1 和 -KD2 细胞中,静息[Ca2+]i 和硫辛加精诱导的[Ca2+]i 的增加都有所降低。在 SphK1-KD1 中测量到的激动剂诱导的[Ca2+]i 增加被削弱。在没有细胞外 Ca2+ 的情况下,硫代甘氨酸诱导的[Ca2+]i 上升迅速下降,表明从细胞质中清除 Ca2+ 的能力增强。与此一致,质膜 Ca2+ ATPase(PMCA)-1 和-4 及其辅助亚基 basigin 均强烈上调。通过特异性激动剂或细胞外 S1P 激活 S1P-GPCR 并不能挽救 SphK1 敲除的影响,这表明 S1P-GPCR 并未参与其中。脂质测量结果表明,在 SphK1-KD2 细胞中,不仅 S1P,而且二氢鞘氨醇、神经酰胺和乳糖基甘油三酯也明显减少。SphK2和S1P裂解酶上调,表明通过鞘脂降解途径的通量增加。最后,组蛋白乙酰化在 SphK1-KD2 细胞中增强,组蛋白去乙酰化酶抑制剂伏立诺他诱导 EA.hy926 细胞中 PMCA1 和 basigin 的 mRNA 和蛋白质水平上调。这些数据首次显示了 S1P 代谢对 PMCA1 和 basigin 的转录调控。结论是在 EA.hy926 细胞中敲除 SphK1 会通过增加组蛋白乙酰化上调 PMCA,从而引起细胞 Ca2+ 平衡的长期改变。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A role for plasma membrane Ca2+ ATPases in regulation of cellular Ca2+ homeostasis by sphingosine kinase-1.

Sphingosine-1-phosphate (S1P) is a ubiquitous lipid mediator, acting via specific G-protein-coupled receptors (GPCR) and intracellularly. Previous work has shown that deletion of S1P lyase caused a chronic elevation of cytosolic [Ca2+]i and enhanced Ca2+ storage in mouse embryonic fibroblasts. Here, we studied the role of sphingosine kinase (SphK)-1 in Ca2+ signaling, using two independently generated EA.hy926 cell lines with stable knockdown of SphK1 (SphK1-KD1/2). Resting [Ca2+]i and thapsigargin-induced [Ca2+]i increases were reduced in both SphK1-KD1 and -KD2 cells. Agonist-induced [Ca2+]i increases, measured in SphK1-KD1, were blunted. In the absence of extracellular Ca2+, thapsigargin-induced [Ca2+]i increases declined rapidly, indicating enhanced removal of Ca2+ from the cytosol. In agreement, plasma membrane Ca2+ ATPase (PMCA)-1 and -4 and their auxiliary subunit, basigin, were strongly upregulated. Activation of S1P-GPCR by specific agonists or extracellular S1P did not rescue the effects of SphK1 knockdown, indicating that S1P-GPCR were not involved. Lipid measurements indicated that not only S1P but also dihydro-sphingosine, ceramides, and lactosylceramides were markedly depleted in SphK1-KD2 cells. SphK2 and S1P lyase were upregulated, suggesting enhanced flux via the sphingolipid degradation pathway. Finally, histone acetylation was enhanced in SphK1-KD2 cells, and the histone deacetylase inhibitor, vorinostat, induced upregulation of PMCA1 and basigin on mRNA and protein levels in EA.hy926 cells. These data show for the first time a transcriptional regulation of PMCA1 and basigin by S1P metabolism. It is concluded that SphK1 knockdown in EA.hy926 cells caused long-term alterations in cellular Ca2+ homeostasis by upregulating PMCA via increased histone acetylation.

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来源期刊
CiteScore
8.80
自引率
2.20%
发文量
121
审稿时长
4-8 weeks
期刊介绍: Pflügers Archiv European Journal of Physiology publishes those results of original research that are seen as advancing the physiological sciences, especially those providing mechanistic insights into physiological functions at the molecular and cellular level, and clearly conveying a physiological message. Submissions are encouraged that deal with the evaluation of molecular and cellular mechanisms of disease, ideally resulting in translational research. Purely descriptive papers covering applied physiology or clinical papers will be excluded. Papers on methodological topics will be considered if they contribute to the development of novel tools for further investigation of (patho)physiological mechanisms.
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