雪莲多糖抑制UVB诱导的炎症反应及调控抗菌肽LL-37的作用研究

Inhibition of UVB-induced Inflammatory Response and Regulation of Antimicrobial Peptide LL-37 by Polysaccharide Extracts from Saussurea laniceps

摘要:

为研究雪莲多糖对UVB诱导角质形成细胞炎症因子表达及抗菌肽LL-37的影响，通过乙醇热回流方法提取雪莲多糖，使用不同浓度的雪莲多糖进行COX-2（炎症通路的关键介质）抑制效果检测。建立UVB照射诱导炎症细胞模型，测定其对前列腺素E2（PGE-2）、TNF-α和IL-1β炎症因子的影响以及雪莲多糖与LL-37表达的关系。采用酶联免疫吸附附实验（ELISA）和蛋白质印迹分析检测炎症因子和LL-37抗菌肽的产生。实验结果表明：雪莲多糖在1 000 μg/mL质量浓度下COX-2抑制率达82.41%，100 μg/mL质量浓度下显著下调HaCaT细胞中PGE-2、TNF-α和IL-1β炎症因子的表达（P<0.01）。此外50、100 μg/mL质量浓度下雪莲多糖增强了LL-37抗菌肽的表达（P<0.01），从而下调TNF-α和IL-1β炎症因子表达以此减轻皮肤炎症。结论：雪莲多糖可以显著抑制由UVB诱导的炎症反应，并且可以通过调控LL-37抗菌肽进一步减缓炎症对皮肤带来的损伤，具有预防UVB照射引起皮肤炎症性损伤的潜力。

Abstract:

In order to study the effects of Saussurea laniceps polysaccharides (SLPs) on the expression of inflammatory factors and antimicrobial peptide LL-37 in UVB-induced keratinocytes, SLPs were extracted by the ethanol thermal reflux method, and SLPs at different concentrations were used to examine the inhibitory effect of COX-2 (a key mediator of inflammatory pathway). A cell model of UVB irradiation-induced inflammation was established to determine the influence of SLPs on prostaglandin E2 (PGE-2), TNF-α and IL-1β inflammatory factors, as well as the relationships of SLPs with LL-37 expression. An enzyme-linked immunosorbent assay (ELISA) and western blot analysis were used to detect the production of inflammatory factors and LL-37 antimicrobial peptide. The results showed that the inhibition rate of COX-2 was 82.41% at 1 000 μg/mL, and the expression of PGE-2, TNF-α and IL-1β inflammatory factors in HaCaT cells was significantly downregulated at 100 μg/mL (P<0.01). In addition, SLPs at 50 μg/mL and 100 μg/mL concentrations enhanced the expression of LL-37 antimicrobial peptide (P<0.01), thereby down-regulating the expression of TNF-α and IL-1β inflammatory factors, then reducing skin inflammation. Conclusion: SLP can significantly inhibit the inflammatory response induced by UVB, and can further slow down the damage caused by inflammation to the skin by regulating LL-37 antimicrobial peptides, which has the potential to prevent skin inflammatory damage caused by UVB irradiation.