Laboratrio de Bioqumica e Microbiologia Aplicada, Departamento de Cincia de Alimentos, ICTA, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brasil.
Cell Biol Int. 2010 Feb 22;34(3):317-23. doi: 10.1042/CBI20090025.
The in vitro cytotoxicity of the antimicrobial peptide P34 was evaluated in different eukaryotic cells. The food-grade bacteriocin nisin was also analysed for comparison. Vero cells were treated with different concentrations (0.02-2.5 microg x ml(-1)) of antimicrobial peptide P34 and nisin. Cell viability and plasma membrane integrity were checked by MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide], NRU (Neutral Red dye uptake) and LDH (lactate dehydrogenase) assays. The EC50 values of the peptide P34 in MTT and NRU assays were 0.60 and 1.25 microg x ml(-1) respectively, while values of nisin found were 0.50 and 1.04 microg x ml(-1). In the LDH assay, the EC50 values were 0.65 and 0.62 microg x ml(-1) for P34 and nisin, respectively. The peptide P34 revealed similar haemolytic activity on human erythrocytes (5.8%) when compared with nisin (4.9%). The effects on viability, motility and acrosomal exocytosis of human sperm were also evaluated. Nisin and P34 showed similar effects on sperm parameters. The evaluation of cytotoxicity of antimicrobial peptides is a critical step to guarantee their safe use.
抗菌肽 P34 的体外细胞毒性在不同的真核细胞中进行了评估。同时也对食品级细菌素乳链菌肽进行了分析比较。用不同浓度（0.02-2.5μg/ml）的抗菌肽 P34 和乳链菌肽处理 Vero 细胞。通过 MTT[3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐]、NRU（中性红染料摄取）和 LDH（乳酸脱氢酶）测定法检查细胞活力和质膜完整性。肽 P34 在 MTT 和 NRU 测定中的 EC50 值分别为 0.60 和 1.25μg/ml，而乳链菌肽的 EC50 值分别为 0.50 和 1.04μg/ml。在 LDH 测定中，P34 和乳链菌肽的 EC50 值分别为 0.65 和 0.62μg/ml。与乳链菌肽（4.9%）相比，P34 对人红细胞的溶血活性相似（5.8%）。还评估了抗菌肽对人精子活力、运动能力和顶体酶外排的影响。乳链菌肽和 P34 对精子参数有相似的影响。抗菌肽细胞毒性的评估是保证其安全使用的关键步骤。
