Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Military Veterinary Institute, Academy of Military Medical Sciences, Academy of Military Sciences, Changchun, 130122, China; Department of Clinical Veterinary Medicine, College of Veterinary Medicine, Jilin University, Changchun, 130062, China.
Department of Clinical Veterinary Medicine, College of Veterinary Medicine, Jilin University, Changchun, 130062, China.
Microb Pathog. 2018 Jun;119:125-130. doi: 10.1016/j.micpath.2018.03.032. Epub 2018 Apr 11.
Currently, accumulating evidence is challenging subtherapeutic therapy. Low-dose Norfloxacin (Nor) has been reported to suppress the immune response and worsen leptospirosis. In this study, we investigated the influence of low-dose Nor (0.03 μg/ml, 0.06 μg/ml, 0.125 μg/ml) on leptospiral gene expression and analyzed the immunomodulatory effects of low-dose Nor-treated leptospires in J774A.1 cells. To study the expression profiles of low-dose Nor-treated leptospires, we chose LipL71/LipL21 as reference genes determined by the geNorm applet in this experiment. The results showed that low-dose Nor up-regulated the expression of FlaB and inhibited the expression of 16S rRNA, LipL32, LipL41, Loa22, KdpA, and KdpB compared with the untreated leptospires. These results indicated that low-dose Nor could regulate leptospiral gene expression. Using RT-PCR, the gene expression of IL-1β and TNF-α in J774A.1 cells was detected. Nor-treated leptospires induced higher expression levels of both IL-1β and TNF-α. However, when analyzed by ELISA, the release of mature IL-1β was reduced compared with that observed in cells induced with no Nor-treated leptospires, although the TNF-α protein level showed no significant change. Our study indicated that the gene expression of leptospires could be modulated by low-dose Nor, which induced less IL-1β release in J774A.1 cells.
目前，越来越多的证据对亚治疗疗法提出了挑战。低剂量诺氟沙星（Nor）已被报道可抑制免疫反应并使钩端螺旋体病恶化。在这项研究中，我们研究了低剂量 Nor（0.03μg/ml、0.06μg/ml、0.125μg/ml）对钩端螺旋体基因表达的影响，并分析了低剂量 Nor 处理的钩端螺旋体在 J774A.1 细胞中的免疫调节作用。为了研究低剂量 Nor 处理的钩端螺旋体的表达谱，我们选择 LipL71/LipL21 作为本实验中 geNorm 应用程序确定的参考基因。结果表明，与未处理的钩端螺旋体相比，低剂量 Nor 上调了 FlaB 的表达，并抑制了 16S rRNA、LipL32、LipL41、Loa22、KdpA 和 KdpB 的表达。这些结果表明，低剂量 Nor 可以调节钩端螺旋体的基因表达。通过 RT-PCR 检测 J774A.1 细胞中 IL-1β 和 TNF-α 的基因表达。Nor 处理的钩端螺旋体诱导了更高水平的 IL-1β 和 TNF-α 的表达。然而，通过 ELISA 分析，与未用 Nor 处理的钩端螺旋体诱导的细胞相比，成熟的 IL-1β 的释放减少，尽管 TNF-α 蛋白水平没有显著变化。我们的研究表明，低剂量 Nor 可以调节钩端螺旋体的基因表达，这导致 J774A.1 细胞中 IL-1β 的释放减少。
