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The effect on dentin-pulp complex by FIP-2 isolated from rat wounded pulp

大鼠损伤牙髓FIP-2对牙本质-牙髓复合体的影响

基本信息

批准号：
17591991
负责人：
ARAI Hideo
金额：
$ 2.24万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591991/
关键词：
dentin-pulp complex rat wounded pulp TNF-α signaling promoter activity transcription factor-binding site over-expression cell death TNF-αパスウェイ

项目摘要

Pulpal wound healing followed by cavity preparation may involve reactionary or reparative dentinogenesis in relation to the cavity position; however, little is known about the molecular responses. We aimed to isolate and analyze genes induced or suppressed in the wounded pulp to identify molecular processes involved in the pulp responses to injury. Twenty-three cDNAs were isolated by cDNA subtraction between healthy and wounded pulp of rats. By library screening, we identified rat 14.7K-interacting protein (rFIP)-2A and B genes homologous to human FIP-2, being involved in regulating membrane trafficking and cellular morphogenesis. RT-PCR analysis showed induction for only rFIP-2B in the wounded pulp. In situ hybridization analysis revealed unique expression of rFIP-2s in adult and embryonic tissues of rats. Transcription of rFIP-2A and B was regulated by alternative use of promoters at rFIP-2 locus. When the rFIP-2A or B-pAcGFP1-Golgi construct was transfected into normal rat kidney (NRK-52E) cells, rFIP-2B was localized in Golgi of whereas rFIP-2A, which is a truncated protein lacking the N-terminal 250 amino acids of rFIP-2B, existed ubiquitously in the cytoplasm. In rat pulp fibroblasts (RPC-C2A) cells, rFIP-2B was significantly induced by tumor necrosis factor (TNF)-α, and the induction was dependent on c-jun N-terminal kinase (JNK) pathway. rFIP-2B was localized in the cytoplasm, and translocated into the nucleus by cell death stimuli. The results suggest that rFIP-2 expression is regulated by the alternative promoter site, and rFIP-2B is a crucial molecule mediated by TNF-a, may be involved in cell death pathway during pulp inflammation.

牙髓伤口愈合后，洞制备可能涉及反应性或修复性牙本质形成的洞的位置，但是，很少有人知道的分子反应。我们的目的是分离和分析基因诱导或抑制在受伤的牙髓，以确定参与牙髓损伤反应的分子过程。采用cDNA差减法从大鼠健康牙髓和创伤牙髓组织中分离出23个cDNA。通过文库筛选，我们发现大鼠14. 7 K相互作用蛋白（rFIP）-2A和B基因与人FIP-2同源，参与细胞膜转运和细胞形态发生的调控。RT-PCR分析显示在受伤的牙髓中仅诱导rFIP-2B。原位杂交分析揭示了rFIP-2在大鼠成年和胚胎组织中的独特表达。通过在rFIP-2位点交替使用启动子调控rFIP-2A和B的转录。当rFIP-2A或B-pAcGFP 1-Golgi构建体转染到正常大鼠肾（NRK-52 E）细胞中时，rFIP-2B定位于Golgi，而rFIP-2A（其为缺失rFIP-2B的N-末端250个氨基酸的截短蛋白）普遍存在于细胞质中。在大鼠牙髓成纤维细胞（RPC-C2 A）中，rFIP-2B可被肿瘤坏死因子（TNF）-α显著诱导，其诱导依赖于c-jun N末端激酶（JNK）途径。rFIP-2B定位于细胞质中，并通过细胞死亡刺激易位到细胞核中。结果提示，rFIP-2的表达受其启动子的调控，rFIP-2B是TNF-α介导的重要分子，可能参与牙髓炎症过程中细胞的死亡途径。