Calcium antagonists ameliorate ischemia-induced endothelial cell permeability by inhibiting protein kinase C

被引:64
作者
Hempel, A
Lindschau, C
Maasch, C
Mahn, M
Bychkov, R
Noll, T
Luft, FC
Haller, H
机构
[1] Humboldt Univ, Franz Volhard Clin, Fac Med Charite, D-13122 Berlin, Germany
[2] Humboldt Univ, Max Delbruck Ctr Mol Med, Fac Med Charite, D-13122 Berlin, Germany
[3] Univ Giessen, Inst Physiol, Giessen, Germany
关键词
cells; ischemia; proteins; calcium;
D O I
10.1161/01.CIR.99.19.2523
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Background-Dihydropyridines block calcium channels; however, they also influence endothelial cells, which do not express calcium channels. We tested the hypothesis that nifedipine can prevent ischemia-induced endothelial permeability increases by inhibiting protein kinase C (PKC) in cultured porcine endothelial cells. Methods and Results-Ischemia was induced by potassium cyanide/deoxyglucose, and permeability was measured by albumin flux. Ion channels were characterized by patch clamp. [Ca2+](i) was measured by fura 2, PKC activity was measured by substrate phosphorylation after cell fractionation. PKC isoforms were assessed by Western blot and confocal microscopy, Nifedipine prevented the ischemia-induced increase in permeability in a dose-dependent manner. Ischemia increased [Ca2+](i), which was not affected by nifedipine. Instead, ischemia-induced PKC translocation was prevented by nifedipine. Phorbol ester also increased endothelial cell permeability, which was dose dependently inhibited by nifedipine. The effects of non- calcium-channel-binding dihydropyridine derivatives were similar. Analysis of the PKC isoforms showed that nifedipine prevented ischemia-induced translocation of PKC-alpha and PKC-zeta. Specific inhibition of PKC isoforms with antisense oligodeoxynucleotides demonstrated a major role for PKC-alpha. Conclusions-Nifedipine exerts a direct effect on endothelial cell permeability that is independent of calcium channels. The inhibition of ischemia-induced permeability by nifedipine seems to be mediated primarily by PKC-alpha inhibition. Anti-ischemic effects of dihydropyridine calcium antagonists could be due in part to their effects on endothelial cell permeability.
引用
收藏
页码:2523 / 2529
页数:7
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