Fujimura T, Shibuya Y, Moriwaki S, Tsukahara K, Kitahara T, Sano T, Nishizawa Y, Takema Y
Biological Science Laboratories, Kao Corporation, Haga, Tochigi, Japan.
Biol Pharm Bull. 2000 Oct;23(10):1180-4. doi: 10.1248/bpb.23.1180.
The fibroblast-populated collagen gel culture method has been evaluated as a dermal model of wound contraction and granulation in tissues during the wound healing process and as an in vitro model of dermal tissue. We previously reported that an extract of Fucus vesiculosus promoted fibroblast-populated collagen gel contraction and that the promotion of the gel contraction was due to the increased expression of integrin alpha2beta1 on the surface of the fibroblasts. In this study, we investigated the active component of the extract of this alga using extraction and fractionation techniques. Water extraction of the alga was followed by precipitation with excess ethanol and then gel filtration with the boundary molecular weight of 30,000. The high molecular weight fraction obtained from gel filtration was fractionated by ion exchange chromatography on diethylaminoethyl cellulose column to give active fractions that have more polar properties. These polar, high molecular weight fractions which contained molecules with fucose and sulfate groups showed significant gel contraction-promoting activity and integrin expression-enhancing activity, and were estimated to be the sulfated-polysaccharide fucoidan. Commercially available fucoidan showed similar activities to the above-described fraction of this alga. Although it remains necessary to precisely identify the specific active component, the above results indicate that fucoidan is the active component which promotes collagen gel contraction, and also indicate the possibility that it dose so by enhancing the integrin alpha2beta1 expression.
成纤维细胞填充胶原凝胶培养方法已被评估为伤口愈合过程中组织伤口收缩和肉芽形成的真皮模型以及真皮组织的体外模型。我们之前报道过，墨角藻提取物可促进成纤维细胞填充胶原凝胶收缩，且凝胶收缩的促进作用归因于成纤维细胞表面整合素α2β1表达的增加。在本研究中，我们使用提取和分级分离技术研究了该藻类提取物的活性成分。藻类经水提取后，用过量乙醇沉淀，然后进行截留分子量为30,000的凝胶过滤。凝胶过滤得到的高分子量级分通过在二乙氨基乙基纤维素柱上进行离子交换色谱分级分离，得到具有更多极性性质的活性级分。这些含有岩藻糖和硫酸根基团分子的极性高分子量级分显示出显著的凝胶收缩促进活性和整合素表达增强活性，据估计为硫酸化多糖岩藻聚糖。市售岩藻聚糖显示出与上述该藻类级分相似的活性。尽管仍有必要精确鉴定特定的活性成分，但上述结果表明岩藻聚糖是促进胶原凝胶收缩的活性成分，并且还表明其可能通过增强整合素α2β1表达来实现这一作用。
