Glycophenotyping of osteoarthritic cartilage and chondrocytes by RT-qPCR, mass spectrometry, histochemistry with plant/human lectins and lectin localization with a glycoprotein.
Glycophenotyping of osteoarthritic cartilage and chondrocytes by RT-qPCR, mass spectrometry, histochemistry with plant/human lectins and lectin localization with a glycoprotein.
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DOI：
10.1186/ar4330
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发表时间：
2013-10-04
影响因子：
4.9
通讯作者：
Gabius HJ
中科院分区：
文献类型：
--
作者：
Toegel S;Bieder D;André S;Altmann F;Walzer SM;Kaltner H;Hofstaetter JG;Windhager R;Gabius HJ
本研究的目的是表征骨关节炎软骨和人软骨细胞的糖表型。从9名骨关节炎（OA）患者中获得膝关节软骨。使用RT-qPCR分析OA软骨细胞中27种糖基转移酶的mRNA水平。此外，使用质谱法定量N-和O-聚糖。在组织学上，从每例患者中选择Mankin评分（MS）≤4或≥9的两个软骨区域，分别代表轻度和重度OA区域。组织切片用（1）一组选定的植物凝集素染色，用于探测OA糖表型，（2）人凝集素半乳糖凝集素-1和-3，和（3）糖蛋白去唾液酸胎球蛋白（ASF）染色，用于可视化β-半乳糖苷特异性内源性凝集素。我们发现OA软骨细胞表达寡甘露糖苷结构以及非、单和二唾液酸化复合型N-聚糖和核心2 O-聚糖。检测到LacdiNAc终止的结构，反映了OA软骨细胞中B4 GALNT 3 mRNA的存在。植物和人类凝集素的染色谱依赖于软骨退化的程度，并且在严重退化的区域中观察到ASF阳性细胞的比率显著较高。总之，OA软骨中糖组的不同方面随着退行性变的进展而改变。特别是，通过半乳糖凝集素-3和泛半乳糖凝集素传感器ASF测量的改变鼓励详细研究半乳糖凝集素在OA中的功能。
This study aimed to characterize the glycophenotype of osteoarthritic cartilage and human chondrocytes. Articular knee cartilage was obtained from nine osteoarthritis (OA) patients. mRNA levels for 27 glycosyltransferases were analyzed in OA chondrocytes using RT-qPCR. Additionally, N- and O-glycans were quantified using mass-spectrometry. Histologically, two cartilage areas with Mankin scores (MS) either ≤4 or ≥9 were selected from each patient representing areas of mild and severe OA, respectively. Tissue sections were stained with (1) a selected panel of plant lectins for probing into the OA glycophenotype, (2) the human lectins galectins-1 and -3, and (3) the glycoprotein asialofetuin (ASF) for visualizing β-galactoside-specific endogenous lectins. We found that OA chondrocytes expressed oligomannosidic structures as well as non-, mono- and disialylated complex-type N-glycans, and core 2 O-glycans. Reflecting B4GALNT3 mRNA presence in OA chondrocytes, LacdiNAc-terminated structures were detected. Staining profiles for plant and human lectins were dependent on the grade of cartilage degeneration, and ASF-positive cells were observed in significantly higher rates in areas of severe degeneration. In summary, distinct aspects of the glycome in OA cartilage are altered with progressing degeneration. In particular, the alterations measured by galectin-3 and the pan-galectin sensor ASF encourage detailed studies of galectin functionality in OA.
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影响因子：
5.2
作者：
Clark MC;Baum LG
通讯作者：
Baum LG
影响因子：
27.4
作者：
Guévremont, M;Martel-Pelletier, J;Reboul, P
通讯作者：
Reboul, P
影响因子：
4.8
作者：
Fiete, Dorothy;Beranek, Mary;Baenziger, Jacques U.
通讯作者：
Baenziger, Jacques U.
影响因子：
4.3
作者：
Gupta, D;Kaltner, H;Brewer, CF
通讯作者：
Brewer, CF
影响因子：
2
作者：
Danguy, A;Rorive, S;Kiss, R
通讯作者：
Kiss, R