α-1,3 岩藻糖基化水平升高可增强 TNFR1 与 TNF-α 的结合能力，进而导致骨关节炎软骨破坏和细胞凋亡。
Elevation of α-1,3 fucosylation promotes the binding ability of TNFR1 to TNF-α and contributes to osteoarthritic cartilage destruction and apoptosis.
机构信息
Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, 229 Taibai North Road, Xi'an, 710069, Shaanxi Province, China.
The Second Clinical Medical College of Shaanxi University of Chinese Medicine, Xianyang, China.
出版信息
Arthritis Res Ther. 2022 Apr 29;24(1):93. doi: 10.1186/s13075-022-02776-z.
BACKGROUND
Osteoarthritis (OA) is the most common form of arthritis and is characterized by the degradation of articular cartilage and inflammation of the synovial membrane. Fucosylation is an important feature of protein N/O-glycosylation and is involved in a variety of pathological processes, including inflammation and cancer. However, whether fucosylation impacts the OA pathological process is unknown.
METHODS
Total proteins were extracted from cartilage samples obtained from patients with OA (n = 11) and OA rabbit models at different time points (n = 12). OA-associated abnormal glycopatterns were evaluated by lectin microarrays and lectin blots. The expression of fucosyltransferases involved in the synthesis of α-1,3 fucosylation was assessed by semi-qPCR. The synthesis of α-1,3 fucosylation mediated by FUT10 was interrupted by the transfection of siRNA, and the effect of α-1,3 fucosylation on OA-associated events was assessed. Then, immunoprecipitation and lectin blotting were used to investigate the relationship between the α-1,3 fucosylation level of tumor necrosis factor receptor superfamily member 1A (TNFR1) and OA. Finally, a TNFR1 antibody microarray was fabricated to evaluate the effect of α-1,3 fucosylation on the ability of TNFR1 to bind to tumor necrosis factor-α (TNF-α).
RESULTS
Elevated α-1,3 fucosylation was observed in cartilage from OA patients, rabbit models, and chondrocytes induced by TNF-α (fold change> 2, p< 0.01). Our results and the GEO database indicated that the overexpression of FUT10 contributed to this alteration. Silencing the expression of FUT10 impaired the ability of TNFR1 to bind to TNF-α, impeded activation of the NF-κB and P38/JNK-MAPK pathways, and eventually retarded extracellular matrix (ECM) degradation, senescence, and apoptosis in chondrocytes exposed to TNF-α.
CONCLUSION
The elevation of α-1,3 fucosylation is not only a characteristic of OA but also impacts the OA pathological process. Our work provides a new positive feedback loop of "inflammation conditions/TNF-α/FUT10/α-1,3 fucosylation of TNFR1/NF-κB and P38/JNK-MAPK pathways/proinflammatory processes" that contributes to ECM degradation and chondrocyte apoptosis.
背景
骨关节炎（OA）是最常见的关节炎形式，其特征在于关节软骨降解和滑膜炎症。岩藻糖基化是蛋白质 N/O-糖基化的一个重要特征，参与多种病理过程，包括炎症和癌症。然而，岩藻糖基化是否影响 OA 病理过程尚不清楚。
方法
从 OA 患者（n=11）和不同时间点的 OA 兔模型（n=12）的软骨样本中提取总蛋白。通过凝集素微阵列和凝集素印迹评估与 OA 相关的异常糖模式。通过半 qPCR 评估参与α-1,3 岩藻糖基化合成的岩藻糖转移酶的表达。通过转染 siRNA 中断 FUT10 介导的α-1,3 岩藻糖基化，评估α-1,3 岩藻糖基化对 OA 相关事件的影响。然后，通过免疫沉淀和凝集素印迹来研究肿瘤坏死因子受体超家族成员 1A（TNFR1）的α-1,3 岩藻糖基化水平与 OA 之间的关系。最后，构建了 TNFR1 抗体微阵列来评估α-1,3 岩藻糖基化对 TNFR1 与肿瘤坏死因子-α（TNF-α）结合能力的影响。
结果
在 OA 患者、兔模型和 TNF-α诱导的软骨细胞中观察到α-1,3 岩藻糖基化水平升高（倍数变化>2，p<0.01）。我们的结果和 GEO 数据库表明，FUT10 的过表达导致了这种改变。沉默 FUT10 的表达会损害 TNFR1 与 TNF-α 的结合能力，阻碍 NF-κB 和 P38/JNK-MAPK 途径的激活，最终减缓 TNF-α 作用下的细胞外基质（ECM）降解、衰老和凋亡。
结论
α-1,3 岩藻糖基化水平的升高不仅是 OA 的特征，而且还影响 OA 的病理过程。我们的工作提供了一个新的正反馈环“炎症条件/TNF-α/FUT10/TNFR1 的α-1,3 岩藻糖基化/NF-κB 和 P38/JNK-MAPK 途径/促炎过程”，有助于 ECM 降解和软骨细胞凋亡。
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