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目的探讨活性氧对原代培养的猪肺动脉内皮细胞钙离子浓度的影响。方法原代培养的猪肺动脉内皮细胞分别用不同浓度的H2O2刺激2 min,用双波长荧光分光光度计观察细胞浆内钙离子浓度的变化;另外,分别以不同时间用10μM的H2O2刺激,观察细胞浆内钙离子浓度的变化。结果 1μM的H2O2即能引起胞内钙离子浓度显著增高,并且H2O2刺激浓度和胞内钙离子浓度的增高有明显量效关系。钙离子浓度由对照组的(110.85±32.33)nM升高至1μM组的(138.44±56.21)nM、10μM组的(171.83±37.92)nM、100μM组的(192.56±34.43)nM,后者与对照组比较,差异均有统计学意义(均P<0.05)。用10μM的H2O2分别刺激内皮细胞在120 s以后可见到胞内钙离子浓度增高,120 s时钙离子浓度为(171.83±37.92)nM,300 s时钙离子浓度为(176.31±43.17)nM,与对照组比较,差异均有统计学意义(均P<0.05)。结论活性氧能够升高原代培养的猪肺动脉内皮细胞的胞内钙浓度,猪肺动脉内皮细胞对H2O2的刺激有一定的耐受性。
Objective To investigate the effects of reactive oxygen species (ROS) on the calcium concentration of primary cultured porcine pulmonary artery endothelial cells. Methods Primary cultured porcine pulmonary artery endothelial cells were stimulated with different concentrations of H2O2 for 2 min. The changes of intracellular calcium concentration were observed by dual-wavelength fluorescence spectrophotometer. In addition, cells were stimulated with 10 μM H2O2 at different times, Changes in plasma calcium concentration. Results 1μM H2O2 can cause a significant increase in intracellular calcium concentration, and H2O2-stimulated concentration and intracellular calcium concentration increased significantly dose-effect relationship. The calcium concentration increased from (110.85 ± 32.33) nM in the control group to (138.44 ± 56.21) nM in the 1μM group, (171.83 ± 37.92) nM in the 10μM group, and (192.56 ± 34.43) nM in the 100μM group, The differences were statistically significant (all P <0.05). After 120 s stimulation with 10 μM H2O2, the intracellular Ca2 + concentration was increased after 120 s, the Ca2 + concentration was (171.83 ± 37.92) nM at 120 s, the Ca2 + concentration was (176.31 ± 43.17) nM at 300 s, and The difference between the two groups was statistically significant (all P <0.05). Conclusion Reactive oxygen species (ROS) can increase the intracellular calcium concentration in cultured primary cultured human pulmonary artery endothelial cells, and that of pulmonary artery endothelial cells in vitro is resistant to H2O2 stimulation.