Regulation of TNFalpha and TNFbeta gene expression in inflammatory bowel disease

炎症性肠病中 TNFα 和 TNFβ 基因表达的调节

• 批准号: 03670345
• 负责人: HIWATASHI Nobuo
• 金额: $ 1.22万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670345/
• 关键词: Inflammatory bowel disease; Crohn's disease; Ulcerative colitis; Cytokines; mRNA; PCR; TNFalpha; TNFbeta; クロ-ン病; γーIFN

Tumor necrosis factor alpha (TNFalpha) and TNFbeta are cytokines with similar immunoregulatory effects binding to their target cells via TNF receptors. To clarify the regulation of TNFalpha and TNFbeta in the mucosal lesions of IBD patients, we examined levels of TNFalpha and TNFbeta mRNA transcripts and these peptide production in the mucosa.METHODS : 1)Intestinal specimens were obtained through an endoscopy or at operation, and total RNA was extracted by a GTP method. RNA was reverse transcribed with M-MLV-RT into cDNA. TNFalpha and TNFbeta mRNA were quantitated using linear PCR methods. As internal controls, cDNA was amplified by PCR with primers for beta-actin, T-cell receptor alpha, ferritin heavy chain. 2)Other mucosa from patients were cultured for 1,2 hours with LPS (10mug/ml) and tRNA in the mucosa was extracted.RESULTS & CONCLUSIONS : The level of TNFalpha mRN was 32-64 fold increased and TNFbeta mRNA was 4-16 fold increased in inflamed mucosa of IBD patients compared to mucosa of control donors. TNFalpha mRNA was 4-8 fold increased and TNFbeta mRNA was not increased in LPS primed IBD mucosa. TNFalpha is an important mediator in response to endotoxin in IBD.

肿瘤坏死因子α（TNF α）和TNF β是具有类似免疫调节作用的细胞因子，通过TNF受体与其靶细胞结合。为了阐明炎症性肠病（IBD）患者肠粘膜病变中TNF α和TNF β的调节作用，我们检测了炎症性肠病（IBD）患者肠粘膜中TNF α和TNF β mRNA转录水平及这些肽的产生。方法：1）通过内窥镜或手术获得肠道标本，用GTP法提取总RNA。用M-MLV-RT将RNA逆转录成cDNA。使用线性PCR方法定量TNF α和TNF β mRNA。作为内部对照，使用β-肌动蛋白、T细胞受体α、铁蛋白重链的引物通过PCR扩增cDNA。2)结果与结论：炎症性肠病患者炎性粘膜中TNF α mRNA水平是正常对照粘膜的32-64倍，TNF β mRNA水平是正常对照粘膜的4-16倍。在LPS致敏的IBD粘膜中，TNF α mRNA增加4-8倍，而TNF β mRNA没有增加。TNF α是IBD中响应内毒素的重要介质。

项目成果

M.Noguchi: "Cytokines and adhesion molecule induces multinncleated giant cells formationi n Crohins disease" gastroenterology. 102. A672- (1992)

M.Noguchi：“细胞因子和粘附分子在克罗辛病中诱导多核巨细胞形成”胃肠病学。

M.NOGUCHI, N.HIWATASHI et al: "Regulation of TNFalpha and TNFbeta gene expression in inflammatory bowel disease." Gastroenterology. 104. (1993)

M.NOGUCHI、N.HIWATASHI 等人：“炎症性肠病中 TNFα 和 TNFβ 基因表达的调节”。

M.Noguchi: "Regulation of TNFα and TNFβ gene expression in inflammatory bowel disease" gastroenterology. 104. (1993)

M.Noguchi：“炎症性肠病中 TNFα 和 TNFβ 基因表达的调节”胃肠病学 104。（1993）

野口 光徳,樋渡 信夫: "炎症性腸疾患(IBD)における腸管局所のTNFα,TNFβ,IFNーγ産生." 日本消化器病学会雑誌. 89. 212 (1992)

Mitsunori Noguchi、Nobuo Hiwatari：“炎症性肠病 (IBD) 肠道内局部产生 TNFα、TNFβ 和 IFN-γ。”日本胃肠病学会杂志 89. 212 (1992)。

野口 光徳、樋渡 信夫、他: "炎症性腸疾患(IBD)における腸管局所のTNFα、TNFβ,IFNγ産生" 日本消化器病学会雑誌. 89. 212 (1992)

Mitsunori Noguchi、Nobuo Hiwatari 等人：“炎症性肠病 (IBD) 肠道中 TNFα、TNFβ 和 IFNγ 的局部产生”日本胃肠病学会杂志 89. 212 (1992)。