Freund Daniel, Brilloff Silke, Ghazy Tamer, Kirschner Stephan, Gäbel Gabor, Hinterseher Irene, Weiss Norbert, Mahlmann Adrian
1 Tissue Engineering Laboratories, Biotechnology Center (BIOTEC), Technische Universität Dresden, Dresden, Germany.
a These authors contributed equally to this paper.
Vasa. 2018 Jun;47(4):295-300. doi: 10.1024/0301-1526/a000700. Epub 2018 Mar 20.
Microarray analysis has been carried out in this pilot study to compare delineated gene expression profiles in the biopsies of skeletal muscle taken from patients with chronic critical limb ischaemia (CLI) and non-ischaemic control subjects.
Biopsy of gastrocnemius muscle was obtained from six patients with unreconstructed CLI referred for surgical major amputation. As control, biopsies of six patients undergoing elective knee arthroplasty without evidence of peripheral arterial occlusive disease were taken. The differences in gene expression associated with angiogenic processes in specimens obtained from ischaemic and non-ischaemic skeletal muscle were confirmed by quantitative real-time polymerase chain reaction (PCR) analysis.
Compared with non-ischaemic skeletal muscle biopsy of chronic-ischaemic skeletal muscle contained 55 significantly up-regulated and 45 down-regulated genes, out of which 64 genes had a known genetic product. Tissue samples of ischaemic muscle were characterized by increased expression of cell survival factors (e. g. tissue factor pathway inhibitor 2) in combination with reduced expression of cell proliferation effectors (e. g. microfibrillar-associated protein 5 and transferrin receptor). The expression of growth factors (e. g. early growth response 3 and chemokine receptor chemokine C-X-C motif ligand 4) which play a central role in arterial and angiogenic processes and anti-angiogenetic factors (e. g. pentraxin 3) were increased in chronic ischaemic skeletal muscle. An increased expression of extracellular matrix proteins (e. g. cysteine-rich angiogenic inducer 61) was also observed.
Gene expression profiles in biopsies of gastrocnemius muscle in patients with chronic critical limb ischaemia showed an increase in pro-survival factors, extracellular matrix protein deposition, and impaired proliferation, compared with non-ischaemic controls. Further studies are required to analyse the endogenous repair mechanism.
在这项初步研究中进行了微阵列分析，以比较慢性严重肢体缺血（CLI）患者和非缺血对照受试者的骨骼肌活检中所描绘的基因表达谱。
从六名因严重肢体缺血而转诊接受大截肢手术的未重建CLI患者获取腓肠肌活检样本。作为对照，采集了六名接受择期膝关节置换术且无外周动脉闭塞性疾病证据的患者的活检样本。通过定量实时聚合酶链反应（PCR）分析证实了缺血和非缺血骨骼肌标本中与血管生成过程相关的基因表达差异。
与非缺血骨骼肌活检相比，慢性缺血骨骼肌包含55个显著上调基因和45个下调基因，其中64个基因具有已知的基因产物。缺血肌肉组织样本的特征是细胞存活因子（如组织因子途径抑制剂2）表达增加，同时细胞增殖效应因子（如微纤维相关蛋白5和转铁蛋白受体）表达降低。在动脉和血管生成过程中起核心作用的生长因子（如早期生长反应3和趋化因子受体趋化因子C-X-C基序配体4）以及抗血管生成因子（如五聚素3）在慢性缺血骨骼肌中的表达增加。还观察到细胞外基质蛋白（如富含半胱氨酸的血管生成诱导因子61）表达增加。
与非缺血对照相比，慢性严重肢体缺血患者腓肠肌活检中的基因表达谱显示促存活因子增加、细胞外基质蛋白沉积以及增殖受损。需要进一步研究来分析内源性修复机制。
