Ito Gen, Kobayashi Takeshi, Takeda Yoshie, Sokabe Masahiro
Department of Physiology, Nagoya University Graduate School of Medicine, Nagoya, Aichi 466-8550, Japan.
Department of Physiology, Nagoya University Graduate School of Medicine, Nagoya, Aichi 466-8550, Japan; Mechanobiology Laboratory, Nagoya University Graduate School of Medicine, Nagoya, Aichi 466-8550, Japan; Mechanobiology Institute Singapore, National University of Singapore, Singapore 117411, Singapore.
Biochem Biophys Res Commun. 2015 Jan 16;456(3):792-8. doi: 10.1016/j.bbrc.2014.12.037. Epub 2014 Dec 13.
Proteoglycans (PGs) are involved in various cellular functions including cell growth, adhesion, and differentiation; however, their physiological roles are not fully understood. In this study, we examined the effect of PG purified from salmon nasal cartilage (SNC-PG) on wound closure using tissue-cultured cell monolayers, an in vitro wound-healing assay. The results indicated that SNC-PG significantly promoted wound closure in NIH/3T3 cell monolayers by stimulating both cell proliferation and cell migration. SNC-PG was effective in concentrations from 0.1 to 10μg/ml, but showed much less effect at higher concentrations (100-1000μg/ml). The effect of SNC-PG was abolished by chondroitinase ABC, indicating that chondroitin sulfates (CSs), a major component of glycosaminoglycans (GAGs) in SNC-PG, are crucial for the SNC-PG effect. Furthermore, chondroitin 6-sulfate (C-6-S), a major CS of SNC-PG GAGs, could partially reproduce the SNC-PG effect and partially inhibit the binding of SNC-PG to cells, suggesting that SNC-PG exerts its effect through an interaction between the GAGs in SNC-PG and the cell surface. Neutralization by anti-CD44 antibodies or CD44 knockdown abolished SNC-PG binding to the cells and the SNC-PG effect on wound closure. These results suggest that interactions between CS-rich GAG-chains of SNC-PG and CD44 on the cell surface are responsible for the SNC-PG effect on wound closure.
蛋白聚糖（PGs）参与多种细胞功能，包括细胞生长、黏附和分化；然而，它们的生理作用尚未完全明确。在本研究中，我们使用组织培养的细胞单层（一种体外伤口愈合检测方法），检测了从鲑鱼鼻软骨中纯化得到的PG（SNC-PG）对伤口闭合的影响。结果表明，SNC-PG通过刺激细胞增殖和细胞迁移，显著促进了NIH/3T3细胞单层的伤口闭合。SNC-PG在0.1至10μg/ml的浓度范围内有效，但在较高浓度（100 - 1000μg/ml）时效果明显减弱。软骨素酶ABC消除了SNC-PG的作用，这表明硫酸软骨素（CSs）作为SNC-PG中糖胺聚糖（GAGs）的主要成分，对SNC-PG的作用至关重要。此外，SNC-PG GAGs的主要CS——硫酸软骨素6-硫酸酯（C-6-S），可以部分重现SNC-PG的作用，并部分抑制SNC-PG与细胞的结合，这表明SNC-PG通过SNC-PG中的GAGs与细胞表面之间的相互作用发挥其作用。抗CD44抗体中和或CD44基因敲低消除了SNC-PG与细胞的结合以及SNC-PG对伤口闭合的作用。这些结果表明，SNC-PG中富含CS的GAG链与细胞表面的CD44之间的相互作用是SNC-PG对伤口闭合起作用的原因。
