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

• 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; 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

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，然后进行空腔制备，可能涉及与腔位置有关的反动或修复性牙本质发生。但是，对分子反应知之甚少。我们的目的是隔离和分析受伤的果肉中诱导或抑制的基因，以鉴定涉及果肉对损伤反应的分子过程。通过cDNA减法在健康和受伤的大鼠果肉之间分离二十三个cDNA。通过库筛选，我们确定了大鼠14.7k相互作用的蛋白（RFIP）-2a和B基因与人类FIP-2同源，并参与调节膜运输和细胞形态发生。 RT-PCR分析显示了受伤的纸浆中仅RFIP-2B的诱导。原位杂交分析显示，RFIP-2在大鼠的成年和胚胎组织中的独特表达。 RFIP-2A和B的转录通过RFIP-2基因座的启动子的替代使用来调节。当RFIP-2A或B-PACGFP1-GOLGI构建体转化为正常大鼠肾脏（NRK-52E）细胞时，RFIP-2B位于GOLGI中，而RFIP-2A的Golgi则是一种截断的蛋白质，该蛋白缺乏N-末端250的250氨基氨基氨基氨基，RFIP-2B的RFIP-2B，存在于Ubipitiply中。在大鼠浆成纤维细胞（RPC-C2A）细胞中，RFIP-2B被肿瘤坏死因子（TNF）-α显着诱导，并且诱导的依赖于C-JUN N末端激酶（JNK）途径。 RFIP-2B位于细胞质中，并通过细胞死亡刺激转化为细胞核。结果表明，RFIP-2表达受替代启动子位点的调节，RFIP-2B是由TNF-A介导的关键分子，可能参与纸浆注射过程中的细胞死亡途径。