Antagonistic effects of cyproheptadine and anisodamine on [Ca2+]i elevation induced by TNFα in endothelial cell strains |
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Authors: | WANG Li-zan ZHANG Qing-zhu ZHU Fan-he LUN Ning |
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Institution: | 1. Department of Pathophysiology, Jining Medical College, Jining 272013, China;
2. Department of Pharmacology, College of Pharmacy, Shandong University, Jinan 250012, China;
3. Department of Biochemistry, Jining Medical College, Jining 272013, China |
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Abstract: | AIM: To study the effects of cyproheptadine (Cyp) and anisodamine (Ani) on the changes of intracellular free Ca2+ concentration (Ca2+]i) induced by tumor necrosis factor (TNFα) in single endothelial cells, and to explore the mechanisms of TNFα 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 Ca2+]i in single endothelial cell was determined by laser scanning confocal microscopy (LSCM). RESULTS: Ca2+]i in single endothelial cell after stimulation of TNFα 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 Ca2+]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 TNFα (1.2×10-9 mol·L-1)-induced Ca2+]i elevation. CONCLUSIONS: TNFα markedly induces elevation of Ca2+]i in single endothelial cell, it may be an important mechanism of TNFα-induced shock and tissue injury. Cyp and Ani obviously suppress TNFα-induced Ca2+]i elevation, which probably is one of the mechanisms of their antishock effects. |
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Keywords: | Cyproheptadine Anisodamine Tumor necrosis factor Endothelium Cells Calcium |
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