Advanced glycation end-products attenuate human mesenchymal stem cells and prevent cognate differentiation into adipose tissue, cartilage, and bone
Advanced glycation end-products attenuate human mesenchymal stem cells and prevent cognate differentiation into adipose tissue, cartilage, and bone
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DOI：
10.1359/jbmr.050514
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发表时间：
2005-09-01
影响因子：
6.2
通讯作者：
Nagata, K
中科院分区：
文献类型：
--
作者：
Kume, S;Kato, S;Nagata, K
晚期糖基化终产物（AGES）在老年和糖尿病患者的各种组织的长寿命蛋白质上积累，并与慢性并发症（包括肌肉骨骼疾病）有关。人间充质干细胞（MSCs）在组织修复过程中可能分化为成熟的肌肉骨骼组织，但AGES对MSCs的发病作用尚不清楚。材料和方法：将牛血清白蛋白与葡萄糖、甘油醛或乙醇醛（分别指定为AGE-1、AGE-2或AGE-3）孵育制备AGES。检测age处理细胞的增殖、凋亡和活性氧（ROS）生成。免疫组织化学和免疫印迹法检测AGE受体（RAGE）表达。使用中和抗RAGE的抗scrum检查RAGE介导的信号的参与。分化为脂肪组织、软骨和骨的形态学和生物化学监测与特定的标记。结果：AGE-2和AGE-3降低了活细胞数量和5-溴-2'脱氧尿苷（BrdU）掺入，增加了细胞内ROS的生成和凋亡细胞的百分比，而对照组的非结合BSA和AGE-1则没有。MSCs表达RAGE， AGE-2和AGE-3刺激RAGE的表达。这些AGES抑制了脂肪生成分化（通过油红O染色、脂蛋白脂肪酶生成和细胞内甘油三酯含量检测）和软骨生成分化（通过红素O染色和II型胶原生成检测）。在成骨分化中，AGE-2和AGE-3增加了碱性磷酸酶活性和细胞内钙含量；然而，von Kossa染色显示矿化丢失和成熟骨结节形成。RAGE抗血清部分阻止age诱导的细胞事件。结论：AGE-2和AGE-3可能通过抑制骨髓间充质干细胞的成熟而导致骨髓间充质干细胞体内质量损失和组织修复延迟。AGE-RAGE相互作用可能参与了age对MSCs的有害作用。
Introduction: Advanced glycation end-products (AGES) are accumulated on long-lived proteins of various tissues in advanced age and diabetes mellitus and have been implicated in chronic complication, including musculoskeletal disorders. Human mesenchymal stem cells (MSCs) potentially differentiate into mature musculoskeletal tissues during tissue repair, but the pathogenetic role of AGES on MSCs is unclear.Materials and Methods: AGES were prepared by incubating BSA with glucose, glyceraldehydes, or glycolaldehyde (designated as AGE-1, AGE-2, or AGE-3, respectively). Proliferation, apoptosis, and reactive oxygen species (ROS) generation were assayed in AGE-treated cells. The expression of the receptor for AGE (RAGE) was examined by inummohistochemistry and Western blotting. Involvement of RAGE-mediated signaling was examined using a neutralizing antiscrum against RAGE. Differentiation into adipose tissue, cartilage, and bone were morphologically and biochemically monitored with specific markers for each.Results: AGE-2 and AGE-3, but not control nonglycated BSA and AGE-1, reduced the viable cell number and 5-bromo-2'deoxyuridine (BrdU) incorporation with increased intracellular ROS generation and the percentage of apoptotic cells. MSCs expressed RAGE and its induction was stimulated by AGE-2 and AGE-3. These AGES inhibited adipogenic differentiation (assayed by oil red O staining, lipoprotein lipase production, and intracellular triglyceride content) and chondrogenic differentiation (assayed by safranin O staining and type II collagen production). On osteogenic differentiation, AGE-2 and AGE-3 increased alkaline phosphatase activity and intracellular calcium content; however, von Kossa staining revealed the loss of mineralization and mature bone nodule formation. The antiserum against RAGE partially prevented AGE-induced cellular events.Conclusion: AGE-2 and AGE-3 may lead to the in vivo loss of MSC mass and the delay of tissue repair by inhibiting the maturation of MSC-derived cells. The AGE-RAGE interaction may be involved in the deleterious effect of AGEs on MSCs.