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首页> 外文期刊>International journal of peptide research and therapeutics >The Effect of Antimicrobial Peptide Temporin-Ra on Cell Viability and Gene Expression of Pro-inflammatory Factors in A549 Cell Line
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The Effect of Antimicrobial Peptide Temporin-Ra on Cell Viability and Gene Expression of Pro-inflammatory Factors in A549 Cell Line

机译：抗菌肽Temporin-Ra对A549细胞系细胞活力和促炎因子基因表达的影响

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摘要

Antimicrobial peptide Temporin-Ra was isolated from the skin secretions of marsh frog Rana ridibunda. Temporin-Ra was chemically synthesized and purified using RP-HPLC technique. The cytotoxicity of peptide on lung airway epithelial cell line (A549) and peripheral blood mononuclear cells (PBMC) was studied by MTT assay. Furthermore, the effect of Temporin-Ra on the expression of pro-inflammatory factors such as IL-1β and IL-8 in A549 cell line was evaluated at peptide concentrations of 15, 30 and 50 mug/mL for 6, 12 and 24 h using semi-quantitative RT-PCR and real-time PCR methods. The result of our experiments revealed that Temporin-Ra decreased cell viability about 3–13 % in A549 cells and 4–6 % in PBMC cells. Moreover, Temporin-Ra induced the mRNA expression of IL-1β and IL-8 genes in a dose- and time-dependent manner. According to our results, maximum IL-8 mRNA expression was observed after a 24-h treatment of cancer cells with 50 mug/mL peptide with approximately 18-fold increase in expression. The least expression level of IL-1beta was observed after 6-h of incubation in the presence of 15 mug/mL peptide with 2.5-fold increase in expression whereas the most expression level was obtained following 24 h-treatment with 50 mug/mL peptide with 26-fold increase in mRNA expression. Eventually, the present study highlights the role of Temporin-Ra as a potent antimicrobial peptide in the activation and maintenance of inflammatory processes.
机译：从沼泽蛙蛙蛙的皮肤分泌物中分离出抗菌肽Temporin-Ra。 Temporin-Ra是通过RP-HPLC技术化学合成和纯化的。 MTT法研究了该肽对肺气道上皮细胞系（A549）和外周血单核细胞（PBMC）的细胞毒性。此外，在肽浓度为15、30和50 mug / mL的情况下，分别在6、12和24小时内评估了Temporin-Ra对A549细胞系中促炎因子如IL-1β和IL-8表达的影响。使用半定量RT-PCR和实时PCR方法。我们的实验结果表明，Temporin-Ra在A549细胞中降低了3-13％的细胞活力，在PBMC细胞中降低了4-6％的细胞活力。此外，Temporin-Ra以剂量和时间依赖性的方式诱导IL-1β和IL-8基因的mRNA表达。根据我们的结果，在用50杯/毫升肽处理癌细胞24小时后，观察到最大的IL-8 mRNA表达，表达增加了约18倍。在存在15杯/毫升肽的情况下孵育6小时后，IL-1beta的表达水平最低，表达增加2.5倍;而用50杯/ mL肽处理24小时后，获得最高的表达水平mRNA表达增加26倍。最终，本研究强调了Temporin-Ra作为有效的抗菌肽在激活和维持炎症过程中的作用。

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