Leucine-rich amelogenin peptide: A candidate signaling molecule during cementogenesis
Leucine-rich amelogenin peptide: A candidate signaling molecule during cementogenesis
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DOI：
10.1902/jop.2004.75.8.1126
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发表时间：
2004-08-01
影响因子：
4.3
通讯作者：
Somerman, MJ
中科院分区：
文献类型：
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作者：
Boabaid, F;Gibson, CW;Somerman, MJ
背景：牙骨质是一种重要的矿化组织，然而，其形成的控制仍然不明确。一种假说是成釉细胞和/或上皮静止细胞分泌的釉基质蛋白/肽有助于通过上皮-间充质相互作用控制牙骨质形成。方法：将小鼠成牙骨质细胞（OCCM-30）暴露于富含亮氨酸的釉原蛋白肽（LRAP）中，观察LRAP对成牙骨质细胞增殖活性的影响，并采用北方印迹法检测Cbfa 1基因的表达。核心结合因子α 1（骨钙素），骨桥蛋白（骨桥蛋白）和OPG的实时逆转录聚合酶链反应（RT-PCR）（骨保护素）;和RANKL（NF-κ B配体的受体激活剂）; 3）使用PKA（THFA）、PKC（PKC）结果：LRAP对细胞增殖没有影响，最多6天，与未处理的细胞相比，在最高剂量下观察到细胞生长减少，9天。LRAP以剂量和时间反应方式下调OCN和上调OPN，并抑制矿物结核形成的能力。与对照组相比，LRAP处理的细胞中OPG的转录物增加，但RANKL mRNA水平不受影响。核心结合因子α（Cbfa）mRNA，组成型表达，不受LRAP。信号通路分析表明参与的MAPK途径，因为除了MAPK抑制剂抑制OPN的表达在LRAP处理cells.Conclusion：亮氨酸丰富的釉原蛋白肽似乎有直接的影响成牙骨质细胞的活动，可能证明显着的发展过程中，以及在牙周组织的再生。
Background: Cementum is a critical mineralized tissue; however, control of its formation remains undefined. One hypothesis is that enamel matrix proteins/peptides secreted by ameloblasts and/or epithelial rest cells contribute to the control of cementum formation via epithelial-mesenchymal interactions. Here, we focused on determining whether or not leucine-rich amelogenin peptide (LRAP), translated from an alternatively spliced amelogenin RNA, altered cementoblast behavior.Methods: Immortalized murine cementoblasts (OCCM-30) were exposed to LRAP and evaluated for: 1) proliferative activity; 2) gene expression using Northern blot for Cbfa1 (core binding factor alpha-1); OCN (osteocalcin), OPN (osteopontin), and real-time reverse transcription-polymerase chain reaction (RT-PCR) for OPG (osteoprotegerin); and RANKL (receptor activator of NF-kappaB ligand); 3) signaling pathway using inhibitors of PKA (THFA), PKC (GF109203X), and MAPK (UO126); and 4) mineralization evaluated by von Kossa and Alizarin-red.Results: LRAP had no effect on cell proliferation up to 6 days, with a decrease in cell growth observed at the highest dose by 9 days versus untreated cells. LRAP down regulated OCN and up regulated OPN in a dose- and time-response fashion, and inhibited the capacity of mineral nodule formation. Transcripts for OPG were increased in LRAP-treated cells compared to control, but RANKL mRNA levels were not affected. Core binding factor alpha (Cbfa) mRNA, expressed constitutively, was not affected by LRAP. Signaling pathway assays suggested involvement of the MAPK pathway, since the addition of the MAPK inhibitor suppressed OPN expression in LRAP-treated cells.Conclusion: Leucine-rich amelogenin peptide appears to have a direct effect on cementoblast activity that may prove significant during development as well as in regeneration of periodontal tissues.