钠/氢交换抑制剂和胃质子/钾ATP酶对人多形核白细胞的细胞体积、细胞内pH值及迁移的影响

Effect of inhibitors of Na+/H+-exchange and gastric H+/K+ ATPase on cell volume, intracellular pH and migration of human polymorphonuclear leucocytes.

作者信息

Ritter M, Schratzberger P, Rossmann H, Wöll E, Seiler K, Seidler U, Reinisch N, Kähler C M, Zwierzina H, Lang H J, Lang F, Paulmichl M, Wiedermann C J

机构信息

Dept. of Internal Medicine, University of Innsbruck, Austria.

出版信息

Br J Pharmacol. 1998 Jun;124(4):627-38. doi: 10.1038/sj.bjp.0701864.

DOI:10.1038/sj.bjp.0701864

PMID:9690853

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1565429/

Abstract

Stimulation of chemotaxis of human polymorphonuclear leucocytes (PMNs) with the chemoattractive peptide fMLP (N-formyl-Met-Leu-Phe) is paralleled by profound morphological and metabolic alterations like changes of intracellular pH (pHi) and cell shape. The present study was performed to investigate the interrelation of cell volume (CV) regulatory ion transport, pHi and migration of fMLP stimulated PMNs. 2. Addition of fMLP to PMNs stimulated directed migration in Boyden chamber assays and was accompanied by rapid initial intracellular acidification and cell swelling. 3. Inhibition of the Na+/H+ exchanger suppressed fMLP stimulated cell migration, accelerated the intracellular acidification and inhibited the fMLP-induced cell swelling. 4. Step omission of extracellular Na+ caused intracellular acidification, which was accelerated by subsequent addition of gastric H+/K+ ATPase inhibitor SCH 28080, or by omission of extracellular K+ ions. In addition Na+ removal caused cell swelling, which was further enhanced by fMLP. 5. H+/K+ATPase inhibitors omeprazole and SCH 28080 inhibited stimulated migration and blunted the fMLP-induced increase in CV. 6. Increasing extracellular osmolarity by addition of mannitol to the extracellular solution caused cell shrinkage followed by regulatory volume increase, partially due to activation of the Na+/H+ exchanger. In fMLP-stimulated cells the CV increase was counteracted by simultaneous addition of mannitol. Under these conditions the fMLP stimulated migration was inhibited. 7. The antibacterial activity of PMNs was not modified by Hoe 694 or omeprazole. 8. Western analysis with a monoclonal anti gastric H+/K+ ATPase beta-subunit antibody detected a glycosylated 35 kD core protein in lysates of mouse and human gastric mucosa as well as in human PMNs. 9. The results indicate that fMLP leads to cell swelling of PMNs due to activation of the Na+/H+ exchanger and a K+-dependent H+-extruding mechanism, presumably an H+/K+ ATPase. Inhibition of these ion transporters suppresses the increase in CV and precludes PMNs from stimulated migration.

摘要

趋化肽fMLP（N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸）刺激人多形核白细胞（PMN）的趋化作用时，会伴随深刻的形态学和代谢改变，如细胞内pH值（pHi）和细胞形状的变化。本研究旨在探讨fMLP刺激的PMN的细胞体积（CV）调节性离子转运、pHi与迁移之间的相互关系。2. 在Boyden小室试验中，向PMN添加fMLP可刺激定向迁移，并伴随着快速的初始细胞内酸化和细胞肿胀。3. 抑制Na⁺/H⁺交换体可抑制fMLP刺激的细胞迁移，加速细胞内酸化，并抑制fMLP诱导的细胞肿胀。4. 逐步去除细胞外Na⁺会导致细胞内酸化，随后添加胃H⁺/K⁺ ATP酶抑制剂SCH 28080或去除细胞外K⁺离子会加速这种酸化。此外，去除Na⁺会导致细胞肿胀，fMLP会进一步加剧这种肿胀。5. H⁺/K⁺ATP酶抑制剂奥美拉唑和SCH 28080抑制刺激的迁移，并减弱fMLP诱导的CV增加。6. 通过向细胞外溶液中添加甘露醇来增加细胞外渗透压会导致细胞收缩，随后是调节性体积增加，部分原因是Na⁺/H⁺交换体的激活。在fMLP刺激的细胞中，同时添加甘露醇可抵消CV的增加。在这些条件下，fMLP刺激的迁移受到抑制。7. Hoe 694或奥美拉唑不会改变PMN的抗菌活性。8. 用单克隆抗胃H⁺/K⁺ ATP酶β亚基抗体进行的蛋白质印迹分析在小鼠和人胃黏膜裂解物以及人PMN中检测到一种糖基化的35 kD核心蛋白。9. 结果表明，fMLP由于激活Na⁺/H⁺交换体和一种K⁺依赖性H⁺排出机制（可能是H⁺/K⁺ ATP酶）而导致PMN细胞肿胀。抑制这些离子转运体可抑制CV的增加，并阻止PMN的刺激迁移。