Articular chondrocytes express the receptor for advanced glycation end products - Potential role in osteoarthritis
Articular chondrocytes express the receptor for advanced glycation end products - Potential role in osteoarthritis
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DOI：
10.1002/art.21199
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发表时间：
2005-08-01
影响因子：
--
通讯作者：
Yan, SD
中科院分区：
文献类型：
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作者：
Loeser, RF;Yammani, R;Yan, SD
Objective.晚期糖基化终末产物受体（receptor for advanced glycation end products，简称AGEs）结合多种配体，包括S100蛋白、高迁移率族蛋白I（high mobility group box chromosomal protein I，HMGB-1）和AGEs，所有这些都存在于关节软骨中。刺激TNF信号传导可导致MAP激酶活化和NF-κ B活性增加。本研究的目的是确定软骨细胞是否表达功能性RAGE。使用正常和骨关节炎（OA）软骨从年轻和年老的猴子和人类，软骨细胞裂解物和人软骨提取物的免疫印迹，和逆转录-聚合酶链反应（RT-PCR）分析的RNA与白细胞介素-1（IL-1）和纤连蛋白片段处理的软骨细胞的免疫组化分析软骨细胞的存在。通过用S100 B和HMGB-1刺激软骨细胞并分析ERK MAP激酶和NF-κ B的活化来评估ERK信号传导。还评估了S100 B和HMGB-I刺激基质金属蛋白酶13（MMP-13）产生的能力。使用生物素标记的S100 B的下拉测定用于证明与RAGE的结合。在猴膝关节软骨和人膝关节和踝关节软骨的切片中检测到β-淀粉样蛋白。在老年猴和人类的软骨中观察到了增加的免疫染色，并且在OA组织中进一步增加。还通过免疫印迹和RT-PCR检测到IL-1 β和纤连蛋白片段，其中发现IL-1 β和纤连蛋白片段刺激IL-1 β表达。用S100 B或HMGB-1刺激软骨细胞增加ERK MAP激酶和NF-κ B的p65亚基的磷酸化，并增加MMP-13的产生。S100 B可与软骨细胞RAGE结合。关节软骨细胞表达功能性软骨素。在OA软骨中观察到的TNF的增加和TNF配体刺激软骨细胞MAP激酶和NF-κ B活性以及刺激MMP-13产生的能力表明软骨细胞TNF信号传导可能在OA中起作用。
Objective. The receptor for advanced glycation end products (RAGE) binds multiple ligands, including S100 proteins, high mobility group box chromosomal protein I (HMGB-1), and AGEs, all of which are present in articular cartilage. Stimulation of RAGE signaling can lead to MAP kinase activation and increased NF-kappa B activity. The objective of the present study was to determine if chondrocytes express functional RAGE.Methods. The presence of chondrocyte RAGE was analyzed by immunohistochemistry using normal and osteoarthritic (OA) cartilage from young and old monkeys and humans, immunoblotting of chondrocyte lysates and human cartilage extracts, and reverse transcription-polymerase chain reaction (RT-PCR) analysis of RNA from chondrocytes treated with interleukin-1 (IL-1) and fibronectin fragments. RAGE signaling was evaluated by stimulating chondrocytes with S100B and HMGB-1 and analyzing for activation of the ERK MAP kinase and NF-kappa B. The ability of S100B and HMGB-I to stimulate matrix metalloproteinase 13 (MMP-13) production was also assessed. A pull-down assay using biotin-labeled S100B was used to demonstrate binding to RAGE.Results. RAGE was detected in sections of monkey knee cartilage and human knee and ankle cartilage. Increased immunostaining for RAGE was noted in cartilage from older adult monkeys and humans and was further increased in OA tissue. RAGE was also detected by immunoblotting and by RT-PCR, where IL-1 beta and fibronectin fragments were found to stimulate RAGE expression. Stimulation of chondrocytes with S100B or HMGB-1 increased phosphorylation of the ERK MAP kinase and the p65 subunit of NF-kappa B and increased the production of MMP-13. This signaling was inhibited in cells pretreated with soluble RAGE, and S100B was shown to bind to chondrocyte RAGE.Conclusion. Articular chondrocytes express functional RAGE. The increase in RAGE noted in OA cartilage and the ability of RAGE ligands to stimulate chondrocyte MAP kinase and NF-kappa B activity and to stimulate MMP-13 production suggests that chondrocyte RAGE signaling could play a role in OA.