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牙龈卟啉单胞菌脂多糖刺激的人中性粒细胞中 supervillin 和血管内皮生长因子的 mRNA 表达升高。

Elevated mRNA expression for supervillin and vascular endothelial growth factor in human neutrophils stimulated with lipopolysaccharide from Porphyromonas gingivalis.

作者信息

Morozumi T, Kubota T, Sugita N, Ohsawa Y, Yamazaki K, Yoshie H

机构信息

Department of Periodontology, Faculty of Dentistry, Niigata University, Japan.

出版信息

J Periodontal Res. 2001 Jun;36(3):160-8. doi: 10.1034/j.1600-0765.2001.360304.x.

Abstract

Differential gene expression was examined in human neutrophils stimulated with lipopolysaccharide from Porphyromonas gingivalis (P. gingivalis-LPS) using RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR). LPS from Escherichia coli (E. coli-LPS) was used as control. More than 200 differently expressed transcripts were found in 8 subjects showing differential regulation at the transcriptional level. Densitometric analyses revealed that 42-100 genes were upregulated, while 53-116 genes were downregulated by P. gingivalis-LPS compared with E. coli-LPS. The results of sequencing identification showed the presence of supervillin (SVIL) and vascular endothelial growth factor (VEGF) genes in the clones which were upregulated by P. gingivalis-LPS. Consequently, semiquantitative analyses proved that the level of mRNA for SVIL and VEGF were significantly higher in P. gingivalis-LPS-stimulated neutrophils than in other bacterial (E. coli, Actinobacillus actinomycetemcomitans, Prevotella intermedia) LPS- or synthetic lipid A-stimulated neutrophils. Our findings suggest that an elevated mRNA expression for SVIL could be associated with impaired function of neutrophils when stimulated by P. gingivalis-LPS. Further, the VEGF mRNA over-expression might be related to the pathogenesis of P. gingivalis-associated periodontitis. The RAP-PCR technique used in this study enabled us to identify a number of P. gingivalis-LPS regulated genes which hitherto have not been reported.

摘要

利用任意引物聚合酶链反应(RAP-PCR)进行RNA指纹分析,检测牙龈卟啉单胞菌脂多糖(P. gingivalis-LPS)刺激的人中性粒细胞中的差异基因表达。以大肠杆菌脂多糖(E. coli-LPS)作为对照。在8名受试者中发现了200多个差异表达的转录本,这些转录本在转录水平上呈现差异调节。密度分析显示,与E. coli-LPS相比,P. gingivalis-LPS上调了42 - 100个基因,而下调了53 - 116个基因。测序鉴定结果表明,在P. gingivalis-LPS上调的克隆中存在 supervillin(SVIL)和血管内皮生长因子(VEGF)基因。因此,半定量分析证明,在P. gingivalis-LPS刺激的中性粒细胞中,SVIL和VEGF的mRNA水平显著高于其他细菌(大肠杆菌、伴放线放线杆菌、中间普氏菌)LPS或合成脂多糖A刺激的中性粒细胞。我们的研究结果表明,当受到P. gingivalis-LPS刺激时,SVIL的mRNA表达升高可能与中性粒细胞功能受损有关。此外,VEGF mRNA的过度表达可能与牙龈卟啉单胞菌相关性牙周炎的发病机制有关。本研究中使用的RAP-PCR技术使我们能够鉴定出许多迄今尚未报道的受P. gingivalis-LPS调控的基因。

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