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目的:探讨结构因素对卡拉胶降解物安全性的影响。方法:以结肠上皮细胞(Caco-2)为研究对象,通过MTT法检测3种类型的卡拉胶(κ,ι和λ型)在3种聚合度范围内的细胞毒,并通过测定活性氧、NO、TNF-α、IL-8的变化来观察样品对细胞的致炎能力。结果:硫酸基团含量最低的κ-卡拉胶降解物具有最强的细胞毒,而硫酸基团含量最高的λ-卡拉胶细胞毒性最弱。其中κ-卡拉胶低聚物在质量浓度高于200μg/mL时的存活率低于30%,并且这种细胞毒与活性氧的爆发呈明显相关性。同时,κ-卡拉胶低聚物具有最强的促进炎症因子分泌的能力,NO、TNF-α、IL-8都出现了明显的增高。另外,聚合度也是影响卡拉胶细胞毒性和促炎性因子分泌的关键,聚合度越低,毒性越高,炎症因子的分泌量越大。结论:低聚卡拉胶,特别是κ-型卡拉胶能够通过活性氧的爆发造成结肠上皮细胞损伤,并引起大量炎症因子产生。该机制可能会导致卡拉胶低聚物损伤结肠组织,引发结肠溃疡。
Objective: To investigate the structural factors on the safety of carrageenan degradation products. Methods: Caco-2 cells were used as experimental materials. Cytotoxicity of three types of carrageenan (κ, ι and λ) in three polymerization degrees was detected by MTT assay. NO, TNF-α, IL-8 changes to observe the sample’s ability to stimulate cells. Results: The kappa-carrageenan degradation product with the lowest content of sulfate group had the strongest cytotoxicity, while the λ-carrageenan with the highest sulfate content had the weakest cytotoxicity. The kappa-carrageenan oligomers showed less than 30% survival at concentrations higher than 200 μg / mL, and this cytotoxicity was significantly correlated with reactive oxygen species (ROS) burst. At the same time, κ-carrageenan oligomers have the strongest ability to promote the secretion of inflammatory cytokines, NO, TNF-α, IL-8 were significantly increased. In addition, the degree of polymerization is also the key factor affecting the carrageenan cytotoxicity and proinflammatory cytokine secretion, the lower the degree of polymerization, the higher the toxicity, the greater the secretion of inflammatory cytokines. CONCLUSION: Oligomeric carrageenan, particularly κ-carrageenan, can damage colonic epithelial cells through the release of reactive oxygen species and cause a large number of inflammatory cytokines. This mechanism may cause carrageenan oligomers to damage colonic tissue and cause colonic ulceration.