Hailer N P, Blaheta R A, Harder S, Scholz M, Encke A, Markus B H
Klinik für Allgemein- und Abdominalchirurgie, Johann Wolfgang Goethe-Universität Frankfurt am Main.
Zentralbl Chir. 1994;119(5):347-54.
AIMS OF THE INVESTIGATION: It is known that Verapamil and other calcium channel blockers possess immunosuppressive properties. It has been suggested that they exert their effects by antagonising protein kinase C (PKC) or calmodulin. Our aim was to investigate whether the adhesion of lymphocytes to allogenic endothelial cells and lymphocyte migration are impaired in the presence of Verapamil. The role of PKC and calmodulin during expression of adhesion molecules, the expression of adhesion molecules under the influence of Verapamil and lymphocyte motility on endothelial cell monolayers were investigated.
Human endothelial cells (EC) were obtained from umbilical chord veins and incubated as dissociate primary cultures. Expression of adhesion molecules on EC was determined by quantitative immunofluorescence using the CytoFluor 2300-system. Studies on lymphocyte motility were performed by phase contrast microscopy using video scope analysis.
Expression of Intercellular Adhesion Molecule 1 (ICAM-1), Vascular Cell Adhesion Molecule (VCAM-1) and Endothelial Leukocyte Adhesion Molecule I (ELAM-1) was dose-dependently reduced by the application of PKC-inhibitor H7. Expression of ICAM-1 and VCAM-1 was also inhibited by the calmodulin-antagonist W7. Surprisingly, neither R- nor S-Verapamil inhibited adhesion molecule expression, we even observed significant enhancement of ELAM-1- and ICAM-1-expression. Nevertheless, lymphocyte motility on allogenic EC monolayers was impaired in the presence of R-verapamil, the enantiomer that is inactive as a calcium channel blocker.
Verapamil reduces lymphocyte motility and is therefore effective in impairing lymphocyte-endothelial cell-interactions. These effects seem to be independent of calcium channel blockade and are probably not due to inhibition of PKC or calmodulin.
研究目的：已知维拉帕米和其他钙通道阻滞剂具有免疫抑制特性。有人提出它们通过拮抗蛋白激酶C（PKC）或钙调蛋白发挥作用。我们的目的是研究在维拉帕米存在的情况下淋巴细胞与同种异体内皮细胞的黏附以及淋巴细胞迁移是否受损。研究了PKC和钙调蛋白在黏附分子表达过程中的作用、维拉帕米影响下黏附分子的表达以及淋巴细胞在内皮细胞单层上的运动性。
从脐静脉获取人内皮细胞（EC），作为解离的原代培养物进行培养。使用CytoFluor 2300系统通过定量免疫荧光法测定EC上黏附分子的表达。通过相差显微镜使用视频显微镜分析进行淋巴细胞运动性研究。
应用PKC抑制剂H7后，细胞间黏附分子1（ICAM-1）、血管细胞黏附分子（VCAM-1）和内皮白细胞黏附分子I（ELAM-1）的表达呈剂量依赖性降低。钙调蛋白拮抗剂W7也抑制了ICAM-1和VCAM-1的表达。令人惊讶的是，R-和S-维拉帕米均未抑制黏附分子的表达，我们甚至观察到ELAM-1和ICAM-1表达显著增强。然而，在R-维拉帕米（作为钙通道阻滞剂无活性的对映体）存在的情况下，淋巴细胞在同种异体EC单层上的运动性受损。
维拉帕米降低淋巴细胞运动性，因此在损害淋巴细胞-内皮细胞相互作用方面有效。这些作用似乎与钙通道阻滞无关，可能不是由于抑制PKC或钙调蛋白所致。
