Pathophysiological role of S100A12 protein via the receptor for advanced glycation end products(EN-RAGE)

S100A12 蛋白通过晚期糖基化终末产物受体 (EN-RAGE) 的病理生理作用

• 批准号: 13671203
• 负责人: KOSAKI Atsushi
• 金额: $ 2.37万
• 依托单位: Kansai Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671203/
• 关键词: S100A12; RAGE; Macrophages; IL-6; PPAR; Diabetes mellitus; チアゾリジン; PPARγ

1.The Regulation of EN-RAGE(S100A12) Gene Expression in Human THP-1 MacrophagesEN-RAGE is a ligand for the receptor for advanced glycation end products(RAGE) and may be involved in the development of diabetic macro-and micro-angiopathy. This study is designed to investigate the regulation of EN-RAGE gene expression in human macrophages. The amounts of EN-RAGE mRNA were measured in cultured human THP-1 macrophages after treatment with various stimuli known to modulate atherosclerosis. First, interleukin-6(IL-6), a proinflammatory cytokine, increased the level of EN-RAGE mRNA by〜2-fold in a time-and a dose-dependent fashion. EN-RAGE protein was detected in the cultured medium and increased significantly by the addition of IL-6. The induction was abolished by pretreatment with the Jak kinase inhibitor and cycloheximide, but not with the MEK kinase inhibitor. Second, pioglitazone, a thiazolidinedione, decreased the level of EN-RAGE mRNA by〜25% of the basal in a time-and a dose-dependent fa … More shion. Pioglitazone also inhibited the induction of EN-RAGE mRNA by IL-6. These results indicate the production of EN-RAGE is induced by IL-6 through de novo protein synthesis via the JAK-STAT kinase pathway and inhibited by the activation of peroxisome proliferator activated receptor-γ(PPARγ) in human macrophages.2.Increased plasma S100A12(EN-RAGE) levels in patients with type 2 diabetesS100A12, also called EN-RAGE or CAAF1, is a ligand for the receptor for advanced glycation end products(RAGE). It has been shown that S100A12 induces adhesion molecules such as VCAM-1 and ICAM-1 in the vascular endothelial cell and mediates migration and activation of monocytes/macrophages through RAGE binding, and that infusion of lipopolysaccharide(LPS) into mice causes time-dependent increase of S100A12 in the plasma. Therefore, circulating S100A12 protein may be involved in chronic inflammation in the atherosclerotic lesion. In this study, we developed an ELISA system which uses specific monoclonal antibodies against recombinant human S100A12 to measure plasma S100A12 levels in patients with diabetes. On using our S100A12 ELISA assay system, concentrations of plasma S100A12 were more than twice as high in the patients with diabetes compared to those without. Using univariate analysis in all subjects, plasma S100A12 concentrations correlated with HbA1c, fasting glucose, high sensitivity CRP and white blood cell count. Step-wise multiple regression analyses, however, revealed that only white blood cell count and HbA1c remained significant independent determinants of plasma S100A12 concentration. These results suggest that plasma S100A12 protein levels are regulated by factors related to subclinical inflammation and glucose control in patients with type 2 diabetes. Less

1.人THP-1巨噬细胞中EN-S100 A12基因表达的调控EN-S100 A12是糖基化终末产物受体的配体，可能参与糖尿病大血管和微血管病变的发生。本研究旨在探讨人巨噬细胞中EN-B1基因表达的调控。在用已知调节动脉粥样硬化的各种刺激物处理后，在培养的人THP-1巨噬细胞中测量EN-BMPmRNA的量。首先，白细胞介素-6（IL-6），一种促炎细胞因子，以时间和剂量依赖的方式使EN-BMPmRNA的水平增加了102倍。在培养液中检测到EN-CRP蛋白，并通过添加IL-6而显著增加。用Jak激酶抑制剂和放线菌酮预处理可消除诱导作用，但用MEK激酶抑制剂则不能。第二，吡格列酮，一种噻唑烷二酮类药物，以时间和剂量依赖性的方式降低EN-binding mRNA水平， ...更多信息 shion。吡格列酮还抑制IL-6诱导的EN-binding mRNA的表达。这些结果表明，EN-β的产生是由IL-6通过JAK-STAT激酶途径通过从头蛋白合成诱导的，并被人巨噬细胞中过氧化物酶体增殖物激活受体-γ（过氧化物酶体增殖物激活受体-γ）的激活抑制。2. 2型糖尿病患者血浆S100 A12（EN-β）水平升高S100 A12，也称为EN-β或CAAF 1，是晚期糖基化终产物受体（AGEs）的配体。研究表明，S100 A12诱导血管内皮细胞中的粘附分子如VCAM-1和ICAM-1，并通过LPS结合介导单核细胞/巨噬细胞的迁移和活化，并且将脂多糖（LPS）输注到小鼠中导致血浆中S100 A12的时间依赖性增加。因此，循环S100 A12蛋白可能参与动脉粥样硬化病变的慢性炎症。在这项研究中，我们开发了一种ELISA系统，该系统使用针对重组人S100 A12的特异性单克隆抗体来测量糖尿病患者血浆S100 A12水平。在使用我们的S100 A12 ELISA检测系统时，糖尿病患者的血浆S100 A12浓度是非糖尿病患者的两倍多。所有受试者中使用单因素分析，血浆S100 A12浓度与HbA 1c、空腹血糖、高敏CRP和白色血细胞计数相关。然而，逐步多元回归分析显示，只有白色血细胞计数和HbA 1c仍然是血浆S100 A12浓度的重要独立决定因素。这些结果表明，血浆S100 A12蛋白水平受2型糖尿病患者亚临床炎症和血糖控制相关因素的调节。少

项目成果

Hasegawa, Takamasa: "The Regulation of EN-RAGE(S100A12) Gene Expression in Human THP-1 Macrophages."Atherosclerosis. 171. 211-218 (2003)

Hasekawa, Takamasa：“人 THP-1 巨噬细胞中 EN-RAGE (S100A12) 基因表达的调节。”动脉粥样硬化。

Hasegawa T., Kosaki A.et al.: "The Regulation of EN-RAGE (S100A12) Gene Expression in Human THP-1 Macrophages"Atherosclerosis. (In press). (2003)

Hasekawa T.、Kosaki A.等人：“人类 THP-1 巨噬细胞中 EN-RAGE (S100A12) 基因表达的调节”动脉粥样硬化。

Hasegawa T, Kosaki A, Kimura T, Matsubara H, Mori Y, Okigaki M, Masaki H, Toyoda N, Inoue-Shibata M, Kimura Y, Nishikawa M, Iwasaka T.: "The Regulation of EN-RAGE(S100A12) Gene Expression in Human THP-1 Macrophages."Atherosclerosis. 171. 211-218 (2003)

Hasekawa T、Kosaki A、Kimura T、Matsubara H、Mori Y、Okigaki M、Masaki H、Toyoda N、Inoue-Shibata M、Kimura Y、Nishikawa M、Iwasaka T.：“EN-RAGE（S100A12）基因的调控