噻庚啶和山莨菪碱拮抗TNF_α诱导的内皮细胞[Ca^(2+)]_i增高  

Antagonistic effects of cyproheptadine and anisodamine on [Ca^(2+)]_i elevation induced by TNF_α in endothelial cell strains

作　　者：王立赞[1] 张庆柱[2] 朱凡河[1] 论宁[3] 

机构地区：[1]济宁医学院病理生理学教研室,山东济宁272013 [2]山东大学药学院药理学教研室,山东济南250012 [3]济宁医学院生物化学教研室,山东济宁272013

出　　处：《中国病理生理杂志》2002年第9期1077-1080,共4页Chinese Journal of Pathophysiology

基　　金：山东省科委资助项目 (No.970 116 )

摘　　要：目的 :研究噻庚啶 (Cyp )和山莨菪碱 (Ani)对肿瘤坏死因子 (TNFα)诱导单个内皮细胞内Ca2 + 浓度([Ca2 + ]i)变化的影响 ,以探TNFα 介导休克和Cyp、Ani的抗休克的机制。方法 :人脐静脉内皮细胞株 (ECV30 4 )接种于 35mm含 2mLDMEM培养基的组织培养盘中培养。Fluo - 3/AM负载细胞 ,激光扫描共聚焦显微技术 (LSCM)测定单个内皮细胞 [Ca2 + ]i。结果 :TNFα 使单个内皮细胞 [Ca2 + ]i 呈剂量依赖性升高 ,在 6 0s内达到峰值 ,然后下降并保持在基础水平之上。共聚焦扫描图像显示细胞核区 [Ca2 + ]i 升高比胞浆区明显 ,下降比胞浆区慢。Cyp (3× 10 -5mol/L或 6× 10 -5mol/L)、Ani (2× 10 -5mol/L或 4× 10 -5mol/L)均能显著抑制由TNFα(1.2× 10 -9mol/L)诱导的单个内皮细胞 [Ca2 + ]i 升高。结论 :TNFα 诱导内皮细胞 [Ca2 + ]i 升高可能是TNFα 介导休克的重要机制 ;Cyp和Ani抑制TNFα诱导的 [Ca2 + ]i 升高可能是其抗休克作用的机制之一。AIM: To study the effects of cyproheptadine (Cyp) and anisodamine (Ani)on the changes of intracellular free Ca 2+ concentration ([Ca 2+ ] i) induced by tumor necrosis factor (TNF α) in single endothelial cells, and to explore the mechanisms of TNF α-mediated shock and antishock actions of Cyp and Ani. METHODS: Human umbilical vein endothelial cell strains(ECV304) were seed in 35 mm tissue culture dish with 2 mL DMEM culture medium. The cultured cells were loaded by Fluo-3/AM. The spatial distribution and the dynamic changes of [Ca 2+ ] i in single endothelial cell was determined by laser scanning confocal microscopy(LSCM). RESULTS: [Ca 2+ ] i in single endothelial cell after stimulation of TNF α rapidly increased in a dose-dependent manner and approached the peak value within 60 seconds, afterwards, decreased and kept above the basal level. The confocal scanning image showed that [Ca 2+ ] i elevation was more obvious in nuclear than in cytoplasma, and decreased slowly. Cyp (3×10 -5 , 6×10 -5 mol/L) and Ani (2×10 -5 , 4×10 -5 mol·L -1 ) markedly inhibited TNF α (1.2×10 -9 mol·L -1 )-induced [Ca 2+ ] i elevation. CONCLUSIONS: TNF α markedly induces elevation of [Ca 2+ ] i in single endothelial cell, it may be an important mechanism of TNF α-induced shock and tissue injury. Cyp and Ani obviously suppress TNF α-induced [Ca 2+ ] i elevation, which probably is one of the mechanisms of their antishock effects.

关 键 词：噻庚啶 山莨菪碱 肿瘤坏死因子 内皮细胞 钙 

分 类 号：R971.92[医药卫生—药品] R331.32[医药卫生—药学]