Department of Biochemical Science and Technology, National Taiwan University, 1 Roosevelt Road, Sec 4, Taipei 10617, Taiwan.
Peptides. 2012 Dec;38(2):197-207. doi: 10.1016/j.peptides.2012.10.003. Epub 2012 Oct 23.
Trichomonas vaginalis is a parasitic protozoan that causes sexually transmitted infections (STIs) worldwide. The infection is dangerous and easily spreads within a community. Also, some cases of drug resistance were reported. Previously, we reported that the shrimp anti-lipopolysaccharide factor (SALF), an antimicrobial peptide of 24 amino acids, modulates inflammatory responses and inhibits T. vaginalis growth. To date, there is no report on the mechanism of SALF's actions in T. vaginalis' adherence to HeLa cells. In this research using an ELISA, we found that the SALF downregulated the release of proinflammatory cytokines (tumor necrosis factor (TNF)-α, interleukin (IL)-1α, IL-6, IL-8, and monocyte chemoattractant protein (MCP)-1) secreted by T. vaginalis which was adhering to HeLa cells. We also performed real-time PCR experiments to determine the roles of the SALF in the expressions of several proinflammatory genes. Through a Western blot analysis, we determined that SALF treatment inhibited T. vaginalis-treated HeLa cells through the p38 and NF-κB pathways. Furthermore, we used different inhibitors to confirm the pathway by ELISA and Western blotting. Taken together, it is apparent that the SALF suppresses T. vaginalis-induced secretion of proinflammatory cytokines by HeLa cells by acting through the p38 and NF-κB pathways.
阴道毛滴虫是一种寄生的原生动物,它会在全球范围内引起性传播感染(STIs)。这种感染很危险,容易在社区内传播。此外,还报告了一些耐药病例。此前,我们报道了虾抗脂多糖因子(SALF),一种 24 个氨基酸的抗菌肽,可以调节炎症反应并抑制阴道毛滴虫的生长。迄今为止,还没有关于 SALF 在阴道毛滴虫黏附 HeLa 细胞中的作用机制的报告。在这项使用 ELISA 的研究中,我们发现 SALF 下调了黏附在 HeLa 细胞上的阴道毛滴虫释放的促炎细胞因子(肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1α、IL-6、IL-8 和单核细胞趋化蛋白(MCP)-1)。我们还进行了实时 PCR 实验,以确定 SALF 在几种促炎基因表达中的作用。通过 Western blot 分析,我们确定 SALF 通过 p38 和 NF-κB 途径抑制了阴道毛滴虫处理的 HeLa 细胞。此外,我们使用不同的抑制剂通过 ELISA 和 Western blot 来验证该途径。综上所述,SALF 通过 p38 和 NF-κB 途径抑制阴道毛滴虫诱导的 HeLa 细胞分泌促炎细胞因子是显而易见的。