该研究旨在探究十二烷基硫酸钠(sodium lauryl sulfate, SLS)胁迫对益生菌外用功效的影响。通过比较自由基清除率和超氧化物歧化酶(superoxide dismutase,SOD)活性筛选具有高抗氧化活性的益生菌,将筛选出的黏膜乳杆菌用1 mmol/L SLS胁迫培养,代谢组学分析差异代谢物判断潜在功效并在细胞内进行进一步功效验证及体外毒理学安全性评估。通过代谢组学分析,鉴定出192种差异代谢物,并发现包括泛酸、甲基补骨脂黄酮等抗氧化应激相关产物在SLS胁迫的黏膜乳杆菌XWL溶胞物(LM+)中含量明显大于未胁迫样品(LM)(P＜0.001)。且证实了LM+对HaCaT细胞毒性和对SLS诱导产生的活性氧(reactive oxygen species, ROS)清除效果优于LM。鸡胚绒毛尿囊膜试验与Ames试验证明两种溶胞物在体积分数40%以下无刺激性和致突变性。综上,黏膜乳杆菌 XWL在SLS胁迫下会分泌更多益生物质来应对胁迫环境,从而加强对SLS导致的HaCaT细胞损伤的针对性保护作用,该研究为益生菌的功效提升提供思路。
To investigate the effect of sodium lauryl sulfate (SLS) stress on the efficacy of probiotics in vitro. Probiotics with high antioxidant activity were screened by comparing the scavenging ability of hydroxyl radicals, and the enzyme activity of superoxide dismutase. The screened Lactobacillus mucosae was cultured under stress of 1 mmol/L SLS, and metabolomic analysis of the differential metabolites was performed to determine the potential efficacy. Furthermore, the cytotoxicity and toxicology were validated by in vitro assay. In the results, a total of 192 differential metabolites were identified by metabolomic analysis, and anti-oxidative stress-related products, including pantothenic acid and methyl psoralen flavonoids, were found to be significantly elevated in L. mucosae XWL after SLS exposure (P＜0.001). The cytotoxicity of SLS-stressed L. mucosae XWL lysate (LM+) was also confirmed to be superior to that of the unstressed sample (LM), and the scavenging of SLS-induced reactive oxygen species (ROS) was significantly enhanced on HaCaT cells. The HET-CAM and Ames assays demonstrated that both lysates were non-irritating and mutagenic below 40% volume fraction. In conclusion, L. mucosae XWL secretes more probiotic substances in response to the stressful environment under SLS stress, thus enhancing the targeted protection against SLS-induced HaCaT cell damage, and this study provides ideas for the enhancement of probiotic efficacy.
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