中文摘要：

目的探讨经典钙拮抗剂维拉帕米（Ver）、硝苯地平（Nif）、地尔硫（Dil）及本课题组合成的新型钙拮抗剂-碘化N-正丁基氟哌啶醇（F2）通过调节钙非依赖磷脂酶A2（iPLA2）抗心脏微血管内皮细胞（CMECs）缺氧/复氧（H/R）损伤的作用与机制。方法培养大鼠原代CMECs,制作H/R模型,在H/R的基础上,用不同浓度梯度的钙拮抗剂及F2处理细胞,比色法测定细胞培养上清液中乳酸脱氢酶（LDH）的漏出反映细胞损伤程度;酶联免疫吸附法（ELISA）测定白介素-6（IL-6）和花生四烯酸（AA）的含量;TUNEL法检测细胞晚期凋亡水平;Real-time PCR检测iPLA2mRNA表达,Western blot检测iPLA2蛋白表达。结果钙拮抗剂F2、Ver和Nif均可量效-依赖性地减少H/R过程中LDH漏出（P〈0.05）,降低IL-6和AA含量（P〈0.05）,减少细胞凋亡（P〈0.05）,而Dil对上述指标均无作用;同时,F2和Ver均可量效-依赖性地降低iPLA2mRNA和蛋白表达水平,Nif和Dil对H/R过程中iPLA2mRNA和蛋白表达水平并无影响。结论H/R可导致CMECs损伤,钙拮抗剂F2、Ver、Nif能保护CMECs抗H/R损伤,Dil无抗H/R损伤的作用;而F2和Ver是部分通过调节iPLA2抗H/R损伤作用的。

英文摘要：

Aim To investigate the effects of classic calcium antagonists verapamil（ Ver）,nifedipine（ Nif）,diltiazem（ Dil） and the novel calcium antagonist N-nbutyl haloperidol iodide（ F2） which was synthesized by our lab by regulating Ca~（2＋）-independent phospholipase A2（ iPLA2） on hypoxia/reoxygenation（ H/R） injury of cardiac microvascular endothelial cells（ CMECs）and the mechanisms. Methods The CMECs were isolated from SD neonatal rats. The H/R model was established,then cells were treated with different concentrations of calcium antagonists and F2. The content of LDH in the cell supernatant was measured by colorimetric method. The levels of IL-6 and AA in cell supernatant were measured by ELISA; and late-stage apoptosis was measured by TUNEL. The mRNA and protein expression levels of iPLA2 in CMECs were examined by real time-PCR and Western blot analysis. Results Calcium antagonists except Dil decreased the generation of LDH,IL-6 and AA in a dose-dependent manner（ P〈0. 05）,and reduced the apoptosis（ P〈0. 05）. F2 and Ver decreased the mRNA and protein expression of iPLA2 in a dose-dependent manner,while there were no such effects for Nif and Dil. Conclusions Calcium antagonists except Dil have protective effects against H/R injury. F2 and Ver protect CMECs against H/R injury partly through iPLA2.