Functional analyais of defense molecule using transgenic mice

使用转基因小鼠进行防御分子的功能分析

• 批准号: 16390369
• 负责人: FUKUSHIMA Kouhei
• 金额: $ 9.09万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390369/
• 关键词: mucosal defense; transgenic mice; epithelial cells; ulcerative colitis; intestinal flora; total proctocolectomy; 遺伝子導入; 炎症性腸疾患; resistin-like molecule; 抗菌タンパク; 大腸

The aims of the present study were to analyze molecular function of resistin-like molecule β (RELM-β) and deleted in malignant brain tumors 1(DMBT-1), which had been identified in colonic epithelial cells by bacterial reconstitution of germ-free mice.We established RELM-β transgenic mice using pCAG vector. After a long time and started to investigate expression of the trannsgene. Its expression in the colon was unexpectedly weak due to unknown reasons. In addition, there was no significant difference between control and TG mice after challenge of dextran sulfate sodium, which are well accepted as an inducer of colitis in mice.In vitro experiments, we found antibacterial effects of RELM-β and still try to confirm this function. Recombinant RELM-β bound to the cell surface of Staphylococcus aureus and destroyed its membrane. This protein is present as dimmer in stools.On the other hand, we struggled to clone full-length DMBT-1 and make its recombinant protein and finally failed. The recent report suggests that cloning of DMBT-1 requires exchange of nucleotides from original to modified sequences (Caroline End, et al. Protein expression and purification 41, 275 (2005).

本研究的目的是分析抵抗素样分子β（RELM-β）和缺失在恶性脑肿瘤1（DMBT-1）中的分子功能，该分子是通过无菌小鼠的细菌重建在结肠上皮细胞中鉴定出的。我们利用pCAG载体建立了RELM-β转基因小鼠。过了很长一段时间，开始研究转基因的表达。由于未知原因，其在结肠中的表达出乎意料地弱。此外，在接受右旋糖酐硫酸钠攻击后，对照小鼠和TG小鼠之间没有显着差异，右旋糖酐硫酸钠被广泛认为是小鼠结肠炎的诱导剂。在体外实验中，我们发现了RELM-β的抗菌作用，并且仍在尝试证实这一功能。重组 RELM-β 与金黄色葡萄球菌的细胞表面结合并破坏其细胞膜。该蛋白在粪便中以二聚体形式存在。另一方面，我们努力克隆全长DMBT-1并制备其重组蛋白，但最终失败了。最近的报告表明，DMBT-1 的克隆需要将核苷酸从原始序列交换为修饰序列（Caroline End, et al. Protein expression andpurification 41, 275 (2005)。

项目成果

A new drug delivery system targeting ileal epithelial cells induced electrogenic sodium absorption. Surgery of the Alimentary Tract San Diego

一种针对回肠上皮细胞的新药物递送系统诱导电化学钠吸收。

• 发表时间: 2006
• 作者: Haneda S、K.Fukushima;et al.
• 通讯作者: et al.

An approach to analyze mechanisms of intestinal adaptation following total proctocolectomy

分析全直肠结肠切除术后肠道适应机制的方法

• 发表时间: 2006
• 期刊: J Gastrointest Surg. 10(5)
• 作者: 福島浩平;et al.
• 通讯作者: et al.

What is unknown in Pouchitis and to where we should go

Pouchitis 中未知的事物以及我们应该去往的地方

• 发表时间: 2006
• 作者: K.Fukushima;et al.
• 通讯作者: et al.

Molecular analysis of colonic transformation in the ileum total colectomy in rats

大鼠回肠全结肠切除术中结肠转化的分子分析

• 发表时间: 2006
• 作者: Tanaka T;et al.;Izumi N;Fukushima K.
• 通讯作者: Fukushima K.

Molecular analysis of colonic transformation in the ileum after total colectomy in rats

大鼠全结肠切除术后回肠结肠转化的分子分析

• 发表时间: 2006
• 期刊: Surgery (In press)
• 作者: Fukushima K;et al.
• 通讯作者: et al.