College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
J Dairy Sci. 2012 Feb;95(2):759-64. doi: 10.3168/jds.2011-4703.
Pheromonicin-SA (Ph-SA) is a newly developed, engineered multidomain peptide that has a bactericidal effect against Staphylococcus aureus. The objective of this study was to characterize innate immune responses by Staph. aureus-stimulated bovine mammary epithelial cells (BMEC) following treatment with Ph-SA. Primary BMEC from one lactating Holstein cow were isolated and exposed to Staph. aureus for 2 h, and then treated with rifampicin or Ph-SA. Total RNA was isolated from BMEC at 0, 2, 6, 12, and 24 h postinfection, and the mRNA expression of selected genes, including toll-like receptor (TLR)2 and TLR4, IL-1β, IL-6, IL-8, tumor necrosis factor α (TNF-α), and lactoferrin, was quantified by real-time PCR. In the rifampicin group, increases in the expression of mRNA for TNF-α, IL-1β, IL-6, IL-8, and lactoferrin were observed at 6 h postinfection and in the expression of mRNA for TLR2 but not for TLR4 at 12 h postinfection. In the Ph-SA group, increases in the mRNA expression of TLR2, TNF-α, IL-1β, IL-6, IL-8, and lactoferrin were observed at 6 h postinfection, and an increase in TLR4 mRNA expression was observed at 24 h postinfection. At 24 h postinfection, the mRNA expression of TLR4, TNF-α, IL-1β, IL-6, IL-8, and lactoferrin was higher in the Ph-SA group than in the rifampicin group. In conclusion, Ph-SA might promote the expression of mRNA for TLR2, TLR4, the pro-inflammatory cytokines IL-1, IL-6, and TNF-α, the chemotactic factor IL-8, and lactoferrin in Staph. aureus-infected BMEC. Moreover, Ph-SA may be of value as an antibiotic in promoting innate immune responses by Staph. aureus-infected bovine mammary epithelial cells.
乳香菌素 - SA(Ph-SA)是一种新开发的、工程化的多结构域肽,对金黄色葡萄球菌具有杀菌作用。本研究的目的是描述金黄色葡萄球菌刺激牛乳腺上皮细胞(BMEC)后,经 Ph-SA 处理后固有免疫反应的特征。从一头泌乳荷斯坦奶牛中分离出原代 BMEC,用金黄色葡萄球菌孵育 2 小时,然后用利福平或 Ph-SA 处理。在感染后 0、2、6、12 和 24 小时从 BMEC 中分离总 RNA,并通过实时 PCR 定量选择基因(包括 TLR2 和 TLR4、IL-1β、IL-6、IL-8、肿瘤坏死因子 α(TNF-α)和乳铁蛋白)的 mRNA 表达。在利福平组中,在感染后 6 小时观察到 TNF-α、IL-1β、IL-6、IL-8 和乳铁蛋白的 mRNA 表达增加,在感染后 12 小时观察到 TLR2 的 mRNA 表达增加,但 TLR4 的 mRNA 表达没有增加。在 Ph-SA 组中,在感染后 6 小时观察到 TLR2、TNF-α、IL-1β、IL-6、IL-8 和乳铁蛋白的 mRNA 表达增加,在感染后 24 小时观察到 TLR4 的 mRNA 表达增加。在感染后 24 小时,Ph-SA 组中 TLR4、TNF-α、IL-1β、IL-6、IL-8 和乳铁蛋白的 mRNA 表达均高于利福平组。综上所述,Ph-SA 可能促进 TLR2、TLR4、促炎细胞因子 IL-1、IL-6 和 TNF-α、趋化因子 IL-8 和乳铁蛋白在金黄色葡萄球菌感染的 BMEC 中的 mRNA 表达。此外,Ph-SA 可能具有作为抗生素的价值,通过促进金黄色葡萄球菌感染的牛乳腺上皮细胞的固有免疫反应。