Alterations of fucosylation in human osteoarthritis cartilage

doi:10.3969/j.issn.2095-4344.0803

Abstract

Abstract:

BACKGROUND: Knee osteoarthritis is characterized as cartilage degeneration and inflammation. However, the mechanism related to abnormal glycosylation modification of the cartilage remains unclear.
OBJECTIVE: To study the abnormal glycosylation modification associated with osteoarthritis.
METHODS: The lectin microarray was employed to investigate the glycan profile of the articular cartilage from 8 patients with knee osteoarthritis and 10 healthy volunteers. The alterations of glycopatterns of osteoarthritic and normal chondrocytes in vitro were evaluated by lectin microarray. In addition, RT-PCR and western blotting were used to analyze the expression levels of related glycosylation enzymes.
RESULTS AND CONCLUSION: The glycopatterns of Fucα-1, 3/6GlcNAc, multivalent Sia and Manα1-6Man which identified by lectins of PSA, LTL, WGA and NPA increased in the osteoarthritic cartilage tissues and chondrocytes compared with the controls. In contrast, high mannose type of N-glycan and bi/tri/tetr-antennary N-glycans which recognized by Con A and PHA-E+L exhibited a decrease in osteoarthritic cartilage tissues and chondrocytes. According to the results of RT-PCR and western blot, glycosyltransferase FUT9 up-regulated in osteoarthritic cartilage tissues and chondrocytes, which may contribute to the alteration of α-1,3-fucosylation in osteoarthritic cartilage. We concluded that the high level of α-1,3-fucosylation may be related with inflammatory microenvironment in osteoarthritic cartilage.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Osteoarthritis, Glycosylation, Inflammation, Tissue Engineering

CLC Number:

R318

Zhao Lian1, Liu Shao1, Li Yi2, Yu Han-jie3, Zhao Hong-mou2, Liang Xiao-jun2, Yang Jie2, Ji Wei-na2. Alterations of fucosylation in human osteoarthritis cartilage[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(20): 3136-3143.

Figures/Tables 4

2.1 参与者数量分析纳入膝骨关节炎患者8例，对照患者10例，全部进入结果分析(见图1)。 2.2 两组基线资料比较见表2。 2.3 凝集素芯片分析骨性关节炎相关异常糖基化研究中，利用凝集素芯片技术比较8例膝关节骨性关节炎患者以及10例正常关节软骨糖蛋白糖链谱，结果显示：①凝集素PSA识别的Fucα-1,6GlcNAc结构，LTL识别的Fucα1-3Galβ1-4GlcNAc结构，BPL、PNA、MAL-I以及MPL识别的Galβ1-3GalNAc结构，SJA和DBA识别的Terminal in GalNAc，WGA识别的Multivalent Sia结构以及NPA识别的Manα1-6Man结构在膝骨关节炎组中的丰度明显高于健康对照组(图2A)；②凝集素STL识别的trimers and tetramers of GlcNAc, core (GlcNAc) of N-glycan结构，ACA识别的T antigen结构，RCA120识别的β-Gal结构，Con A识别的high-Mannose结构以及PHA-E+L识别的Bisecting GlcNAc, bi-antennary N-glycans, tri-and tetra- antennary complex-type N-glycan糖链结构在健康对照组中的丰度明显高于膝骨关节炎组(图2B)；③HCE聚类分析结果显示，膝骨关节炎组与健康对照组之间糖蛋白糖链的种类和丰度存在差异，Fucα-1,3/6GlcNAc岩藻糖基化以及较短的O-聚糖Galβ1-3GalNAc糖链结构在膝骨关节炎组中的丰度较高，而在健康对照组中，STL，Con A以及PHA-E+L识别的N-聚糖结构明显高于膝骨关节炎组(图2C)。软骨组织之间差异凝集素识别的糖链结构以及差异倍数见表3。另外，分别分离膝关节骨性关节炎患者软骨以及正常关节软骨进行体外培养，待细胞融合度达到80%左右时提取总蛋白，利用凝集素芯片比较体外培养的关节软骨细胞糖蛋白糖链谱的差异。结果显示，①凝集素PSA识别的Fucα-1,6GlcNAc结构，LTL识别的Fucα1-3Galβ1- 4GlcNAc结构，RCA120识别的β-Gal结构，NPA识别的Manα1-6Man结构，BS-I识别的α-Gal结构以及WGA识别的Multivalent Sia结构在骨性关节炎软骨细胞中的丰度明显高于正常软骨细胞(图3A)；②在正常软骨细胞中，凝集素PWM识别的(GlcNAc)n结构，EEL识别的Galα1-3 (Fucα1-2)Gal结构，Con A识别的high-Mannose结构，PHA-E+L识别的Bisecting GlcNAc, bi-antennary N-glycans, tri-and tetra-antennary complex-type N- glycan结构以及UEA-I识别的Fucα1-2Galβ1-4Glc(NAc)这些糖链结构的丰度明显高于骨性关节炎软骨细胞(图3B)；③HCE聚类分析结果显示骨性关节炎软骨细胞与正常软骨细胞在糖链分布上也存在差异(图3C)。软骨细胞之间差异凝集素识别的糖链结构以及差异倍数见表4。"

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