Dudek Slawomir, Ziółko Ewa, Kimsa-Dudek Magdalena, Solarz Krzysztof, Mazurek Urszula, Wierzgoń Aleksander, Kokot Teresa, Muc-Wierzgoń Małgorzata
Department of Pharmacognosy and Phytochemistry, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Sosnowiec, Poland.
Department of Internal Medicine, School of Public Health, Silesian Medical University in Katowice, Bytom, Poland.
Arch Immunol Ther Exp (Warsz). 2017 Apr;65(2):175-182. doi: 10.1007/s00005-016-0416-8. Epub 2016 Sep 7.
The similarity of Lyme borreliosis to other diseases and its complex pathogenesis present diagnostic and therapeutic difficulties. The changes that occur at the cellular and molecular levels after a Borrelia sp. infection still remain poorly understood. Therefore, the present study focused on the expression of TLR and TLR-signaling genes in human dermal fibroblasts in the differentiation of an infection with Borrelia burgdorferi sensu lato spirochetes. Normal human dermal fibroblasts were cultured with the spirochetes of Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii. Total RNA was extracted from the cells using TRIzol reagent. The analysis of the expression profiles of TLRs and TLR-related genes was performed using commercially available oligonucleotide microarrays of HG-U133A. The GeneSpring 12.0 platform and significance analysis of microarrays were used for the statistical analysis of microarray data. The analyses using the oligonucleotide microarray and QRT-PCR techniques permitted to identify the genes encoding TLR4 and TLR6 as specific for infection with B. afzelii and B. burgdorferi sensu stricto. In turn, TLR3 was only characteristic for an infection with B. burgdorferi sensu stricto. There were no changes in the TLR gene expression after infection with B. garinii. Our findings confirm that Borrelia has a major effect on fibroblast gene expression. Further characterization of changes in gene expression may lead to valuable insights into the role of the toll-like receptor in the pathogenesis of Lyme disease and may provide guidelines for the development of diagnostic markers for an infection with a particular Borrelia genospecies. Moreover, this will help to identify better treatment strategies for Lyme disease.
莱姆病与其他疾病的相似性及其复杂的发病机制给诊断和治疗带来了困难。伯氏疏螺旋体感染后在细胞和分子水平上发生的变化仍知之甚少。因此，本研究聚焦于人类皮肤成纤维细胞中Toll样受体（TLR）和TLR信号基因在伯氏疏螺旋体广义种螺旋体感染分化过程中的表达。将正常人皮肤成纤维细胞与狭义伯氏疏螺旋体、阿氏疏螺旋体和伽氏疏螺旋体的螺旋体共同培养。使用TRIzol试剂从细胞中提取总RNA。使用市售的HG-U133A寡核苷酸微阵列对TLR和TLR相关基因的表达谱进行分析。使用GeneSpring 12.0平台和微阵列显著性分析对微阵列数据进行统计分析。使用寡核苷酸微阵列和定量逆转录聚合酶链反应（QRT-PCR）技术的分析允许鉴定编码TLR4和TLR6的基因是阿氏疏螺旋体和狭义伯氏疏螺旋体感染所特有的。反过来，TLR3只是狭义伯氏疏螺旋体感染所特有的。伽氏疏螺旋体感染后TLR基因表达没有变化。我们的研究结果证实伯氏疏螺旋体对成纤维细胞基因表达有重大影响。基因表达变化的进一步表征可能会对Toll样受体在莱姆病发病机制中的作用提供有价值的见解，并可能为开发特定伯氏疏螺旋体基因种感染的诊断标志物提供指导。此外，这将有助于确定更好的莱姆病治疗策略。
