通过激光介导的显微切割分析关节炎滑膜中的血管基因表达。

Analysis of vascular gene expression in arthritic synovium by laser-mediated microdissection.

作者信息

Hashimoto Atsushi, Tarner Ingo H, Bohle Rainer M, Gaumann Andreas, Manetti Mirko, Distler Oliver, Steinmeyer Jürgen, Ulfgren Ann-Kristin, Schulz Andreas, Gay Steffen, Müller-Ladner Ulf, Neumann Elena

机构信息

Department of Medicine and Rheumatology, Justus-Liebig-University of Giessen, Giessen, and University Hospital Regensburg, Germany.

出版信息

Arthritis Rheum. 2007 Apr;56(4):1094-105. doi: 10.1002/art.22450.

DOI:10.1002/art.22450

PMID:17393418

Abstract

OBJECTIVE

In rheumatoid arthritis (RA), formation of new blood vessels is necessary to meet the nutritional and oxygen requirements of actively proliferating synovial tissue. The aim of this study was to analyze the specific synovial vascular expression profiles of several angiogenesis-related genes as well as CD82 in RA compared with osteoarthritis (OA), using laser-mediated microdissection (LMM).

METHODS

LMM and subsequent real-time polymerase chain reaction were used in combination with immunohistochemical analysis for area-specific analysis of messenger RNA (mRNA) and protein expression of vascular endothelial growth factor (VEGF), VEGF receptor 1 (VEGFR-1), VEGFR-2, hypoxia-inducible factor 1alpha (HIF-1alpha), HIF-2alpha, platelet-derived growth factor receptor alpha (PDGFRalpha), PDGFRbeta, inhibitor of DNA binding/differentiation 2 (Id2), and CD82 in RA and OA synovial microvasculature and synovial lining.

RESULTS

Expression of Id2 mRNA was significantly lower in RA synovial vessels compared with OA synovial vessels (P=0.0011), whereas expression of VEGFR-1 was significantly higher in RA (P=0.0433). No differences were observed for the other parameters. At the protein level, no statistically significant differences were observed for any parameter, although Id2 levels were 2.5-fold lower in RA (P=0.0952). However, the number of synovial blood vessels and the number of VEGFR-2-expressing blood vessels were significantly higher in RA compared with OA.

CONCLUSION

Our results underscore the importance of area-specific gene expression analysis in studying the pathogenesis of RA and support LMM as a robust tool for this purpose. Of note, our results indicate that previously described differences between RA and OA in the expression of angiogenic molecules are attributable to higher total numbers of synovial and vascular cells expressing these molecules in RA rather than higher expression levels in the individual cells.

摘要

目的

在类风湿关节炎（RA）中，新血管形成对于满足活跃增殖的滑膜组织的营养和氧气需求是必要的。本研究的目的是使用激光介导显微切割（LMM）分析与骨关节炎（OA）相比，RA中几种血管生成相关基因以及CD82的特定滑膜血管表达谱。

方法

LMM及随后的实时聚合酶链反应与免疫组织化学分析相结合，用于对RA和OA滑膜微血管及滑膜衬里中血管内皮生长因子（VEGF）、VEGF受体1（VEGFR-1）、VEGFR-2、缺氧诱导因子1α（HIF-1α）、HIF-2α、血小板衍生生长因子受体α（PDGFRα）、PDGFRβ、DNA结合/分化抑制因子2（Id2）和CD82的信使核糖核酸（mRNA）和蛋白质表达进行区域特异性分析。

结果

与OA滑膜血管相比，RA滑膜血管中Id2 mRNA的表达显著降低（P=0.0011），而RA中VEGFR-1的表达显著更高（P=0.0433）。其他参数未观察到差异。在蛋白质水平上，尽管RA中Id2水平低2.5倍（P=0.0952），但任何参数均未观察到统计学上的显著差异。然而，与OA相比，RA中滑膜血管数量和表达VEGFR-2的血管数量显著更高。