Sialylation regulates myofibroblast differentiation of human skin fibroblasts.
Sialylation regulates myofibroblast differentiation of human skin fibroblasts.
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DOI：
10.1186/s13287-017-0534-1
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发表时间：
2017-04-18
影响因子：
7.5
通讯作者：
Toyoda M
中科院分区：
文献类型：
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作者：
Sasaki N;Itakura Y;Toyoda M
成纤维细胞是维持皮肤稳态和协调生理组织修复和皮肤再生的关键角色。随着衰老和衰老过程发生的成纤维细胞功能障碍导致在老年人中观察到的延迟愈合。衰老和衰老过程中导致成纤维细胞功能障碍的分子机制尚未阐明。以前，在成纤维细胞中观察到糖基化模式的变化在老化和衰老过程中，但这些变化对成纤维细胞功能的影响还没有得到很好的记录。在这里，我们研究了衰老过程中糖基化的变化是否可能对成纤维细胞产生功能性影响。通过使用凝集素的荧光激活细胞分选分析，在早期传代（EP）和晚期传代（LP）皮肤成纤维细胞中检查衰老过程中皮肤成纤维细胞上细胞表面聚糖的变化。通过实时聚合酶链反应或蛋白质印迹分析检查细胞表面聚糖变化的贡献者。在EP成纤维细胞中检测糖基化变化对转化生长因子（TGF）-β1刺激诱导的增殖、迁移、细胞衰老诱导和肌成纤维细胞分化的影响。通过GalNAc-α-O-苄基或唾液酸酶处理进行糖基化的变化。在LP成纤维细胞中观察到糖蛋白的唾液酸化减少和唾液酸酶NEU 1增加。唾液酸化的减少对增殖、迁移或诱导细胞衰老没有任何影响。另一方面，肌成纤维细胞分化受到唾液酸化减少的抑制，表明唾液酸化对于肌成纤维细胞分化是重要的。CD 44在肌成纤维细胞分化所需的脂筏中的定位被唾液酸化的减少抑制。此外，通过唾液酸酶抑制剂恢复LP成纤维细胞中减少的肌成纤维细胞分化。在衰老过程中，随着唾液酸酶的增加，CD 44的去唾液酸化减少了TGF-β1刺激后CD 44在脂筏中的定位，导致肌成纤维细胞分化的抑制。因此，唾液酸化的调节可能是预防和再生治疗与年龄相关的皮肤疾病、美容皮肤改变以及老年人愈合延迟引起的慢性伤口的一种有吸引力的策略。本文的在线版本（doi：10.1186/s13287-017-0534-1）包含补充材料，可供授权用户使用。
Fibroblasts are key players in maintaining skin homeostasis and in orchestrating physiological tissue repair and skin regeneration. Dysfunctions in fibroblasts that occur with aging and the senescent process lead to the delayed healing observed in elderly people. The molecular mechanisms leading to fibroblast dysfunction during aging and the senescent process have not yet been clarified. Previously, changes in patterns of glycosylation were observed in fibroblasts in aging and the senescent process, but the effect of these changes on the function of fibroblasts has not been well documented. Here, we investigated whether changes in glycosylation during the process to senescence may have functional effects on fibroblasts. The changes in cell surface glycans on skin fibroblasts during the process to senescence were examined in early-passage (EP) and late-passage (LP) skin fibroblasts by fluorescence-activated cell sorting analysis using lectins. The contributors to the changes in cell surface glycans were examined by real-time polymerase chain reaction or Western blot analysis. The effects of changes in glycosylation on proliferation, migration, induction of cellular senescence, and myofibroblast differentiation induced by transforming growth factor (TGF)-β1 stimulation were examined in EP fibroblasts. The changes in glycosylation were performed by GalNAc-α-O-benzyl or sialidase treatment. A decrease in sialylation of glycoproteins and an increase in sialidase NEU1 were observed in LP fibroblasts. The reduction of sialylation did not have any effect on proliferation, migration, or induction of cellular senescence. On the other hand, myofibroblast differentiation was inhibited by the reduction of sialylation, indicating that sialylation is important for myofibroblast differentiation. The localization of CD44 in lipid rafts, which is required for myofibroblast differentiation, was inhibited by the reduction of sialylation. Furthermore, reduced myofibroblast differentiation in LP fibroblasts was restored by a sialidase inhibitor. Desialylation of CD44 with increased sialidase during the process to senescence reduced the localization of CD44 in lipid rafts after TGF-β1 stimulation, leading to the inhibition of myofibroblast differentiation. Thus, regulation of sialylation may be an attractive strategy for the prevention and regenerative therapy of age-related skin diseases, cosmetic skin alterations, and chronic wounds caused by delayed healing in elderly people. The online version of this article (doi:10.1186/s13287-017-0534-1) contains supplementary material, which is available to authorized users.