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.

牙髓伤口愈合后进行的空洞准备可能涉及与空洞位置相关的反应性或修复性牙本质发生；然而，我们的目的是分离和分析受伤牙髓中诱导或抑制的基因，以确定参与其中的分子过程。通过对健康和受伤大鼠牙髓的 cDNA 消减，我们分离出 23 个 cDNA。通过文库筛选，我们鉴定了大鼠 14.7K 相互作用蛋白 (rFIP)-2A 和 B 基因同源。人 FIP-2，参与调节膜运输和细胞形态发生。RT-PCR 分析显示，在受伤的牙髓中仅诱导 rFIP-2B。原位杂交分析显示，rFIP-2 在成年大鼠和胚胎组织中具有独特的表达。当 rFIP-2A 或 B-pAcGFP1-Golgi 构建体转染正常大鼠时，rFIP-2A 和 B 的转录受到 rFIP-2 基因座启动子的交替使用的调节。在肾 (NRK-52E) 细胞中，rFIP-2B 定位于高尔基体，而 rFIP-2A 是一种缺少 rFIP-2B N 端 250 个氨基酸的截短蛋白，普遍存在于大鼠牙髓成纤维细胞的细胞质中。 RPC-C2A）细胞中，rFIP-2B 被肿瘤坏死因子（TNF）-α 显着诱导，且诱导依赖于c-jun N 末端激酶 (JNK) 通路定位于细胞质，并通过细胞刺激死亡转位至细胞核。结果表明，rFIP-2 的表达受到替代启动子位点的调节。 2B是TNF-a介导的关键分子，可能参与牙髓炎症过程中的细胞死亡途径。