Elucidation of aging mechanism of hyaluronan knock-down mouse and its application to the evaluation for effect of anti-aging reagents

透明质酸敲低小鼠衰老机制的阐明及其在抗衰老试剂效果评价中的应用

• 批准号: 20300225
• 负责人: KON Atsushi
• 金额: $ 12.06万
• 依托单位: Aomori University of Health and Welfare
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20300225/
• 关键词: ヒアルロン酸; 糖鎖; アンチエイジング; 老化; 発現制御

Hyaluronan(HA), a non-sulfated glycosaminoglycan of high molecular mass, presents in various tissues as a major component of the extracellular matrix and plays an important role in regeneration, wound healing, and anti-aging of organs. We have previously reported that 4-methylumbelliferone(MU) specifically inhibits HA synthesis. In this study, we first investigated the expression of HA-related gene, such as HA synthase(HAS-1, HAS-2, HAS-3), hyaluronidase(Hyal-1, Hyal-2, Hyal-3, Hyal-4), and HA receptor(CD44, RHAMM). We have demonstrated that HAS-2 gene transcription in both epidermis and dermis, and RHAMM transcription in epidermis were reduced in HA-knock-down mice. These data suggested that MU treatment specifically downregulates gene expressions of HAS-2 and RHAMM induced the HA production, resulting in skin aging, such as wrinkle formation and skin dehydration in HA knockdown mice. Next, we have detected sixty one kinds of HA knock-down mice specific genes by using DNA microarray a … More nalysis. Specifically, toll-like receptor 3 gene was strongly down-regulated in HA knock-down mice, suggesting induction of inflammation resulted in skin aging characterize by wrinkle formation. Oncogene expressions such as of tyrosine phosphatase, cyclin Y, and Rho were also down-regulated in HA-knock down mice. Our previous studies have demonstrated MU suppressed the metastasis of malignant tumor cells by inhibiting of HA production. Therefore, these results suggested that down-regulation of these oncogene expression, at least in part, were involved in inhibition of metastasis. On the other hand, gene expression of stromelysin 3, depredating enzyme of extracellular matrix components such as proteoglycan and collagen, strongly up-regulated. These data suggested that the degradation of extracellular matrix resulted in wrinkle formation of HA knock-down mice. Finally, we have administered HA and its oligosaccharides with various molecular weight. As the result, only intradermal injection of high molecular HA decreased skin aging, whereas no change was observed in aged skin by orally or topically administration. Also, skin aging was not reduced by introduction of aging-related gene which was down-regulated in HA knock-down mice, such as COL5A1, COL7A1, RHAMM, toll-like receptor 3, suggesting alone administration of each gene was not effective and cocktail therapy, the combination of multiple genes, was necessary of treatment of skin aging in HA knock-down mice Less

透明质酸（HA）是一种高分子量的非硫酸化糖胺聚糖，作为细胞外基质的主要成分存在于各种组织中，在器官再生、伤口愈合和抗衰老中起重要作用。我们以前曾报道，4-甲基伞形酮（MU）特异性抑制HA的合成。在本研究中，我们首先研究了HA相关基因的表达，如HA合成酶（HAS-1，HAS-2，HAS-3），透明质酸酶（Hyal-1，Hyal-2，Hyal-3，Hyal-4）和HA受体（CD 44，RHAMM）。我们已经证明，在HA敲低小鼠中，表皮和真皮中的HAS-2基因转录以及表皮中的RHAMM转录减少。这些数据表明，MU处理特异性下调HAS-2和RHAMM诱导的HA产生的基因表达，导致皮肤老化，如在HA敲低小鼠中皱纹形成和皮肤脱水。其次，我们利用基因芯片技术检测了61种HA基因敲除小鼠的特异性基因， ...更多信息 分析。具体而言，Toll样受体3基因在HA敲除小鼠中强烈下调，表明炎症诱导导致以皱纹形成为特征的皮肤老化。HA基因敲除小鼠的酪氨酸磷酸酶、细胞周期蛋白Y和Rho等癌基因表达也下调。我们前期的研究表明MU通过抑制HA的产生而抑制恶性肿瘤细胞的转移。因此，这些结果表明，这些癌基因表达的下调，至少部分，参与了抑制转移。另一方面，降解细胞外基质成分如蛋白多糖和胶原的基质分解酶3的基因表达强烈上调。这些数据表明，细胞外基质的降解导致皱纹形成的HA敲低小鼠。最后，我们给药了HA及其各种分子量的寡糖。结果表明，只有皮内注射高分子HA才能降低皮肤老化，而口服或局部给药对老化皮肤无明显影响。此外，通过引入在HA敲除小鼠中下调的衰老相关基因，如COL 5A 1、COL 7A 1、RHAMM、toll样受体3，皮肤衰老没有减少，这表明单独施用每种基因是无效的，并且鸡尾酒疗法，多个基因的组合，是治疗HA敲除小鼠皮肤衰老所必需的。

项目成果

Novel proteoglycan glycotechnology: chemoenzymatic synthesis of chondroitin sulfate-containing molecules and its application

新型蛋白聚糖糖技术：含硫酸软骨素分子的化学酶法合成及其应用

• 发表时间: 2009
• 期刊: Glycoconjugate Journal
• 影响因子: 3
• 作者: M. Yamaguchi;K. Takagaki;K. Kojima;Naohiro Hayashi;Fengchao Chen;I. Kakizaki;A. Kon;M. Endo
• 通讯作者: M. Endo

Skin aging and extracellular matrix.

皮肤老化与细胞外基质。

• 发表时间: 2009
• 作者: Yoshioka;T.;et al.;Kon A
• 通讯作者: Kon A

4-メチルウンベリフェロンによるヒアルロン酸の合成抑制機構

4-甲基伞形酮抑制透明质酸合成的机制

• 发表时间: 2009
• 作者: 今淳;数馬恒平;渡部朋子;渡部一郎;五十嵐愛美;高垣啓一
• 通讯作者: 高垣啓一

Hyaluronan knockdown mice: application to analysis of hyaluronan-mediated pathophysiology

透明质酸敲低小鼠：应用于分析透明质酸介导的病理生理学

• 发表时间: 2008
• 期刊: Inflammation Regenerat 28
• 作者: Okuyama R;et al.;Kon A
• 通讯作者: Kon A

Hyaluronan in the skin and its correlation with dermatopathology

皮肤中的透明质酸及其与皮肤病理学的相关性

• 发表时间: 2011
• 期刊: Trends in Glycosci Glycotechnol
• 作者: Naito;T.;Goto;K.;Morioka;S.;Matsuba;Y.;Akema;T.;Sugiura;T.;Ohira;Y.;Beppu;M.;Yoshioka;T.;Kon A
• 通讯作者: Kon A