Formation of hyaluronic-acid-deficient extracellular matrix induced by an inhibitor for hyaluronic acid synthesis

透明质酸合成抑制剂诱导缺乏透明质酸的细胞外基质的形成

• 批准号: 07680668
• 负责人: NAKAMURA Toshiya
• 金额: $ 0.58万
• 依托单位: Hirosaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07680668/
• 关键词: Hyaluronic acid; Inhibitor; Extracellular matrix; Human skin fibroblasts; 4-Methylumbelliferone; Decorin

Cultivation of human skin fibroblasts in the presence of 0.5 mM 4-methylumbelliferone repressed hyaluronan production of the cell and reduced its chain length, resulting establishment of hyaluronan-deficient extracellular matrix. Cell-free experiments suggested that 4-methylumbelliferone may repress the expression of hyaluronan synthase on the cell surface. Electron-microscopic analysis of the hyaluronan-deficient extracellular matrix showed decrease of extracellular matrix component morphologically. Analysis of hyaluronan-binding proteins in medium showed the decrease of the amount of hyaluronan binding proteins, especially 83kDa protein, in the medium of the cells cultured in the presence of 4-methylumballiferone. Synthesis of proteoglycans of the cells were not affected by 4-methylumbelliferone signifcantly except for that of decorin. The synthesis of decorin was stimulated by 4-methylumbelliferone and, moreover, the number of glycosaminoglycan chains of decorin was increased from 1 to 2 by 4-methylumbelliferone.

在0.5 mM 4-甲基伞形酮存在下培养人皮肤成纤维细胞抑制了细胞的透明质酸产生并减少了其链长，从而建立了透明质酸缺乏的细胞外基质。无细胞实验表明，4-甲基伞形酮可抑制细胞表面透明质酸合成酶的表达。透明质酸缺乏的细胞外基质的电子显微镜分析显示细胞外基质成分的形态减少。对培养液中透明质酸结合蛋白的分析表明，在4-甲基伞形酮存在下培养的细胞的培养液中，透明质酸结合蛋白，特别是83 kDa蛋白的量减少。4-甲基伞形酮对细胞蛋白多糖的合成无明显影响，但对核心蛋白聚糖的合成有影响。4-甲基伞形酮刺激核心蛋白聚糖的合成，而且，4-甲基伞形酮使核心蛋白聚糖的糖胺聚糖链的数目从1增加到2。

项目成果

Shigeru Shibata: "HNK-1-reactive novel oligosaccharide,sulfate-O-3GlcAβ1-4Xy1β1-(4-methyl-umbelliferone),synthesized by cultured human skin fibroblasts" Journal of Biological Chemistry. 270. 13794-13798 (1995)

Shigeru Shibata：“HNK-1-反应性新型寡糖，硫酸盐-O-3GlcAβ1-4Xy1β1-(4-甲基-umbelliferone)，由培养的人皮肤成纤维细胞合成”《生物化学杂志》270。13794-13798 (1995)。

Toshiya Nakamura: "New hyaluronan-degrading enzymes" Connective Tissue. 28. 15-19 (1996)

Toshiya Nakamura：“新型透明质酸降解酶”结缔组织。

Shigeru Shibata: "HNK-reactive novel oligosaccharide,sulfate-O-3GlcA β1-4Xyl β1-(4methylunbelliferone),synthesized by cultured human skin fibroblasts" Journal of Biological Chemistry. 270. 13794-13798 (1995)

Shigeru Shibata：“HNK 反应性新型寡糖，硫酸盐-O-3GlcA β1-4Xyl β1-(4methylunbelliferone)，由培养的人皮肤成纤维细胞合成”《生物化学杂志》270。13794-13798 (1995)。

Kanji Tanaka: "Regulation of hyaluronate metabolism by progesterone in cultured fibroblasts from the human uterine cervix" FEBS Letters. 402. 223-226 (1997)

Kanji Tanaka：“黄体酮对人宫颈培养成纤维细胞中透明质酸代谢的调节”FEBS Letters。