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目的 研究刺尾鱼毒素 (MTX)的细胞毒性及钙通道阻断剂的拮抗效应 ,为探讨可能的防治途径提供依据。方法 采用噻唑蓝 (MTT)比色法检测MTX对细胞的毒性效应及荧光分光光度法测定胞浆内游离Ca2 +的浓度。结果 MTX对LLC PK1细胞的毒性效应有良好的剂量和时间效应关系。MTX 8ng/ml作用 3h已呈现明显的毒性效应 ,随着MTX剂量的增加 ,细胞存活率明显降低 ,与对照组相比差异有极显著性。随着MTX剂量的增加和作用时间的延长其毒性逐渐增大。Ca2 +通道拮抗剂维拉帕米 5× 10 -5mol/L、硝苯吡啶 1× 10 -4mol/L能明显拮抗MTX所致LLC PK1细胞的Ca2 +内流 ;维拉帕米和硝苯吡啶 1× 10 -5mol/L、5× 10 -5mol/L、1× 10 -4mol/L均能够明显降低MTX引起的细胞毒性死亡 ,提高细胞的存活率。结论 MTX的细胞毒性可能是由于细胞内Ca2 +浓度升高引起 ,Ca2 +通道拮抗剂对MTX引起的细胞损伤有一定的保护作用
Objective To study the cytotoxicity of the toxins (MTX) and the antagonism of calcium channel blockers in order to provide evidence for possible prevention and treatment. Methods Toxicity of MTX to cells was measured by MTT assay and the concentration of free Ca2 + in cytoplasm was determined by fluorescence spectrophotometry. Results MTX had a good dose-and time-dependent effect on the cytotoxicity of LLC PK1 cells. MTX 8ng / ml for 3h has shown significant toxic effects, with the MTX dose increased cell viability was significantly reduced, compared with the control group, the difference was significant. With the MTX dose increased and the role of time increased its toxicity gradually increased. Ca 2+ channel antagonist verapamil 5 × 10 -5 mol / L and nifedipine 1 × 10 -4 mol / L significantly antagonized the Ca 2+ influx in LLC PK1 cells induced by MTX. Verapamil and nifedipine 1 × 10 -5 mol / L, 5 × 10 -5 mol / L and 1 × 10 -4 mol / L all could significantly reduce the cytotoxic death caused by MTX and increase the cell survival rate. CONCLUSION: The cytotoxicity of MTX may be due to the increase of intracellular Ca2 + concentration. Ca2 + channel antagonists may have a protective effect on MTX induced cell injury