硝苯地平通过调控COX-2拮抗心脏微血管内皮细胞缺氧/复氧损伤  

Nifedipine antagonizes hypoxia/reoxygenation injuryof cardiac microvascular endothelial cells by regulating COX-2

作　　者：陈润济 柳满 汪彬 李海燕 蔡文锋 石刚刚 CHEN Run-ji;LIU Man;WANG Bin;LI Hai-yan;CAI Wen-feng;SHI Gang-gang(Dept of Pharmacology,Shantou University Medical College,Shantou Guangdong 515041,China)

机构地区：[1]汕头大学医学院药理学教研室,广东汕头515041

出　　处：《中国药理学通报》2020年第7期946-952,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No.81870276)。

摘　　要：目的研究硝苯地平(Nif)对心脏微血管内皮细胞(CMECs)缺氧/复氧(hypoxia/reoxygenation,H/R)损伤的作用与机制。方法通过建立心脏微血管内皮细胞H/R模型;在H/R模型基础上,分别用不同浓度的硝苯地平(Nif)处理内皮细胞,采用比色法测定细胞培养上清液中乳酸脱氢酶(LDH)的漏出情况;采用Annexin V-FITC/PI凋亡检测试剂盒检测细胞的早期凋亡;TUNEL法观察细胞晚期凋亡水平;酶联免疫吸附法(ELISA)测定白介素1β(IL-1β)的含量、内皮细胞黏附分子(ICAM-1)的含量和COX-2产物(PGE2)的含量;Western blot检测COX-2蛋白的表达;结果与Control组相比,COX-2蛋白表达上调、乳酸脱氢酶LDH的漏出增加,产物PGE2的量上升。Nif(10.0、1.0、0.1μmol·L^-1)3个剂量组可呈现量效依赖性地降低炎症因子IL-1β的含量,减轻凋亡,降低H/R损伤COX-2蛋白的高表达、减少产物PGE2的产生。结论 Nif能保护CMECs抗H/R损伤,其机制可能是通过调节COX-2来发挥保护作用。Aim To study the effect of nifedipine on hypoxia/reoxygenation(H/R)injury of cardiac microvascular endothelial cells(CMECs)and its mechanisms.Methods The H/R model of cardiac microvascular endothelial cells was established.On the basis of the H/R model,the endothelial cells were treated with different concentrations of nifedipine,and the leakage of LDH in the supernatant of cultured cells after H/R was measured by colorimetry.The early apoptosis of cells was detected by Annexin V-FITC/PI apoptosis detection kit,and the late apoptosis level was observed by TUNEL method.IL-1β,ICAM-1 and COX-2 product(PGE 2)were detected by enzyme linked immunosorbent assay(ELISA).The expression of COX-2 protein was detected by Western blot.Results Compared with Control group,the expression of COX-2 protein was up-regulated,LDH leakage increased,and PGE 2 increased.Nif(10.0,1.0,0.1μmol·L^-1)couldreduce the content of IL-1β,apoptosis,COX-2 protein expression and PGE 2 production in a dose-dependent manner.Conclusion Nif can protect CMECs against H/R injury,and its protective mechanism may be related tothe regulation of COX-2.

关 键 词：心脏微血管内皮细胞 硝苯地平 COX-2 缺氧/复氧 内皮细胞黏附分子 炎症 

分 类 号：R-332[医药卫生] R322.11