Effects of nonenzymatic glycosylation of extracellular matrix components on cell survival and sensory neurite extension in cell culture
Effects of nonenzymatic glycosylation of extracellular matrix components on cell survival and sensory neurite extension in cell culture
复制标题
DOI：
10.1007/s004150200033
复制
发表时间：
2002-04-01
影响因子：
6
通讯作者：
Thomas, PK
中科院分区：
文献类型：
--
作者：
Luo, ZJ;King, RHM;Thomas, PK
糖尿病感觉性多发性神经病是以垂死型远端轴索病变为特征。它伴随着轴突再生的失败，其中可能涉及细胞外基质的非酶糖基化(糖基化)。本研究在组织培养中研究了基底膜的主要成分IV型胶原和层粘连蛋白的糖基化对神经元存活和轴突生长的影响。层粘连蛋白的快速糖基化是通过与乙醇醛的孵育实现的，IV型胶原的糖基化通过与葡萄糖的孵育实现的。用荧光分析法检测糖基化程度。用糖化或非糖化的层粘连蛋白或IV型胶原作为培养新生大鼠背根神经节(DRG)神经元的底物。对培养物中贴壁细胞、存活细胞和长出神经突起的细胞比例进行评估。IV型胶原糖化2周后细胞贴壁率和轴突形成比例明显减少，但仅糖化4周或5周后细胞存活受影响。与未糖化的层粘连蛋白相比，糖化的层粘连蛋白的3个参数均显著降低。在扫描电子显微镜下，层粘连蛋白和IV型胶原的糖基化在培养的神经元中产生了显著的形态差异。在非糖化底物上培养的患有链脲佐菌素诱导的糖尿病的成年动物的分离的DRG神经元存活得较差，产生的神经突起也较少。因此，IV型胶原和层粘连蛋白的糖基化影响神经元的存活、轴突的产生和细胞的形态，而糖尿病既影响感觉神经元在培养中的存活，也影响它们延伸轴突的能力。
Diabetic sensory polyneuropathy is characterized by a distal axonopathy of dying-back type. It is accompanied by a failure of axonal regeneration, in which nonenzymatic glycosylation (glycation) of the extracellular matrix may be involved. In the present study, the effects of glycation of collagen IV and laminin, major components of basal lamina, on neuron survival and neurite extension were investigated in tissue culture. Fast glycation of laminin was achieved by incubation with glycolaldehyde and glycation of collagen IV by incubation with glucose. The degree of glycation was estimated by fluorescence analysis. Glycated or nonglycated laminin or collagen IV were used as substrates for culture of dorsal root ganglion (DRG) neurons from neonatal rats. Cultures were assessed for the proportion of cells attaching to the substrate, surviving and bearing neurites. Cell attachment and the proportion bearing neurites were significantly reduced on collagen IV glycated for 2 weeks, but survival was only affected by glycation for 4 or 5 weeks. All 3 parameters were significantly reduced on glycated compared with unglycated laminin. Glycation of both laminin and collagen IV produced considerable morphological differences in the cultured neurons on scanning electron microscopy. Dissociated DRG neurons from adult animals with streptozotocin-induced diabetes cultured on nonglycated substrates survived less well and produced fewer neurites. Glycation of collagen IV and laminin thus affects neuronal survival, neurite production and cell morphology, and diabetes affects both the survival of sensory neurons in culture and their ability to extend neurites.