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目的探讨海藻硫酸多糖(sulfated polysaccharide,SP)及其与Fe3+反应生成海藻多糖铁络合物(seaweed sulfated polysaccharide iron complex,SPIC)对氧化应激THP-1细胞的作用。方法用化学检测法比较SP和SPIC主要成分。在此基础上,用分光光度法检测SPIC中铁离子的离解;用电子自旋共振(electron spin resonance,ESR)自旋捕捉技术对络合前后物质的羟自由基清除作用进行对比;用ESR自旋捕捉技术检测氧化应激细胞呼吸爆发的变化;用MTT(methyl thiazolyl tetrazolium,MTT)法观察络合反应前后对正常THP-1细胞和对氧化应激THP-1细胞的存活率的影响。结果 SP与Fe3+反应形成络合物SPIC后,主要作用成分有较大变化,主要是Fe3+浓度含量显著升高,硫酸根和蛋白含量明显下降,而总多糖变化较小。Fe3+离解实验表明4h内SPIC中的铁能全部溶出;ESR自旋捕捉技术表明,络合前后对羟自由基清除作用与浓度密切相关;与SP相比,在相同浓度时SPIC对氧化应激后的细胞存活更高些。结论 SPIC络合上Fe3+后,在一定浓度时对氧化应激细胞的保护作用更佳,主要与络合后的离子性质和浓度密切相关。
Objective To investigate the effects of sulfated polysaccharide (SP) and its reaction with Fe3+ to produce seaweed sulfated iron complex (SPIC) on oxidative stress in THP-1 cells. Methods Chemical analysis was used to compare the main components of SP and SPIC. On this basis, the dissociation of iron ions in SPIC was detected by spectrophotometry; the hydroxyl radical scavenging effects of the complexes before and after complexation were compared by electron spin resonance (ESR) spin trapping technique; the ESR spin was used. Capture technique was used to detect the change of respiratory burst in oxidative stress cells; MTT assay was used to observe the effects of complex reaction on the survival rate of normal THP-1 cells and oxidative stress THP-1 cells. Results After reacting with Fe3+ to form SPIC complexes, the main components of the complexes changed greatly. The main content of Fe3+ was significantly increased, and the contents of sulfate and protein were significantly decreased, while the changes of total polysaccharides were smaller. The Fe3+ dissociation experiments showed that the iron in SPIC was completely dissolved in 4 h. The ESR spin trapping technique showed that the scavenging effect of hydroxyl radicals before and after complexation was closely related to the concentration; compared with SP, SPIC was at the same concentration after oxidative stress. The cell’s survival is higher. Conclusion After SPIC is complexed with Fe3+, the protective effect of SPIC on oxidative stress cells is better at a certain concentration, which is closely related to the nature and concentration of complexed ions.