本研究主要在探討透明質酸 ( Hyaluronan，簡稱HA )促進纖維母細
胞 (fibroblast)爬行能力之主要機制。本實驗首先在二維細胞培養下，比
較出含不同量HA 之膠原蛋白基質表面對纖維母細胞進行爬行之影響差
異 , 實驗結果證實纖維母細胞在含有3-20 µg/cm2 HA 之膠原蛋白基質
上 , 其爬行能力受到顯著促進 , 並以含有5 µg/cm2 HA 實驗組效果最
為顯著。觀察細胞外型與細胞骨架可以發現 , 附著於5 µg/cm2 HA 膠原
蛋白基質上之細胞 , 其在外型與actin cytoskeleton 分布上呈現較為極化
(polarization)現象 , 而lamellipodia 形成情況較為明顯 , 而觀察細胞附
著結構亦發現 , focal adhesion complex 在分布上明顯較集中於
lamellipodia 前緣位置 , 顯示細胞受到環境HA 的刺激後 , 其細胞骨架
組裝與細胞附著方式已有所改變。配合對細胞與膠原蛋白作用之主要分
子β1 integrin 進行觀察可以發現 , β1 integrin 的分布同樣會因為HA的存
在大量分布於lamellipodia 前緣位置 , 顯示HA 的存在促進細胞於
lamellipodia 前緣重新形成附著結構 , 而此一結果可能與細胞爬行能力
受到促進有關。針對HA 之重要受器如CD44 及RHAMM 之作用進行偵
測 , 並對已知相關訊息傳遞路徑之重要蛋白如 FAK , PI3K , ERK1/2 等
進行活性分析結果顯示 , 附著於3-5 µg/cm2 HA 膠原蛋白基質上之細
胞 , 與CD44 相關蛋白如FAK 與PI3K 之活化受到明顯促進 , 而
ERK1/2 之磷酸化則沒有顯著改變 , 顯示以CD44 透過ERM protein 調控
actin cytoskeleton 的機制可能主導細胞骨架組裝與細胞爬行能力受到促
進之結果 , 而RHAMM 透過ERK1/2 調控microtubule 之機制則未被發現 。

　　In this study we investigate the mechanism for hyaluronic acid (HA)
effects on cell migration. The first goal of this study is to compare the
motility of fibroblast on HA-collagen matrix, in which it coated with
various amount of HA. The experimental result demonstrate that fibroblast
have enhanced motility on 3-10 µg/cm2 HA coated collagen matrix;
however, optimum enhancement of migration was seen on 5 µg/cm2 HA
coated collagen matrix. Several differents could be found in fibroblast
which growth on 5 µg/cm2 HA coated collagen matrix. First, Both cell
morphology and actin cytoskeleton became more polarized in them.
Second, lamellipodia formation became observed in leading edge. Third,
cell have numerous focal adhesion complex in lamellipodia. This result
suggest that cell cytoskeleton and focal adhesion was altered after
environmental induction of HA. We also found that the distribution of β1
integrin was localize in leading egde of lamellipodia, indicating
regeneration of β1 integrin-mediated focal adhesion in lamellipodia was
induced by HA. It might be related to enhanced cell motility. By analysis of
the specific receptors of HA, like CD44 and RHAMM, and of
phosphorylation of specific proteins which have been characterized with
HA induction. These proteins include FAK, ERK1/2, PI3K et al. Experental
results show that CD44 related proteins, like FAK and PI3K, were
activated in fibroblast which growth on 5 µg/cm2 HA coated collagen
matrix. However, ERK1/2 were not activated in this condition. Our results
suggest that HA on collagen matrix promotes cell motility by regulate FAK
activity and by ERM protein mediated-actin cytoskeleton regulation. But
not by RHAMM mediated ERK1/2 signaling pathway.

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