Low-dose Norfloxacin-treated leptospires induce less IL-1β release in J774A.1 cells following discrepant leptospiral gene expression
Low-dose Norfloxacin-treated leptospires induce less IL-1β release in J774A.1 cells following discrepant leptospiral gene expression
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DOI：
10.1016/j.micpath.2018.03.032
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发表时间：
2018-06-01
影响因子：
3.8
通讯作者：
Tu, Changchun
中科院分区：
文献类型：
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作者：
Cao, Yongguo;Xie, Xufeng;Tu, Changchun
目前，越来越多的证据正在挑战亚治疗疗法。低剂量诺氟沙星（Nor）已被报道抑制免疫反应和加重钩端螺旋体病。本研究观察了低剂量Nor（0.03 μ g/ml、0.06 μ g/ml、0.125 μ g/ml）对钩体基因表达的影响，并分析了低剂量Nor处理的钩体对J774A.1细胞的免疫调节作用。为了研究小剂量去甲肾上腺素（NOR）处理后钩端螺旋体的表达谱，本实验选择LipL 71/LipL 21作为参考基因，用geNorm applet软件进行检测。结果表明，与未处理的钩端螺旋体相比，低剂量Nor可上调FlaB的表达，抑制16 SrRNA、LipL 32、LipL 41、Loa 22、KdpA和KdpB的表达。结果表明，低剂量Nor对钩体基因表达具有调控作用。RT-PCR检测J774A.1细胞中IL-1 β和TNF-α的基因表达。经去甲肾上腺素处理的钩端螺旋体诱导IL-1 β和TNF-α表达水平升高。然而，当通过ELISA分析时，成熟IL-1 β的释放与未用Nor-treated钩端螺旋体诱导的细胞中观察到的相比减少，尽管TNF-α蛋白水平未显示出显著变化。我们的研究表明，钩端螺旋体的基因表达可以通过低剂量的去甲肾上腺素调节，从而诱导J774A.1细胞释放较少的IL-1 β。
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 mu g/ml, 0.06 mu g/ml, 0.125 mu g/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-1 beta and TNF-alpha in J774A.1 cells was detected. Nor-treated leptospires induced higher expression levels of both IL-1 beta and TNF-alpha. However, when analyzed by ELISA, the release of mature IL-1 beta was reduced compared with that observed in cells induced with no Nor-treated leptospires, although the TNF-alpha 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-1 beta release in J774A.1 cells.