探討人類珍珠素蛋白多醣第五片 段促進神經纖維生長之作用

基底膜 (basement membrane, BM) 是由第四型膠原蛋白 (type IV collagen)

、層黏蛋 白 (laminin) 及蛋白多醣 (proteoglycan) 等物質所構成，主要功能是 扮演維持上皮及內皮細胞型態與細胞附著及做為細胞生長支架 (scaffold) 的功 能。硫酸肝素蛋白多醣 (HSPG) 為 BM 中重要的蛋白多醣類。先前研究指出 H SPG 具有促進神經纖維生長的功能，而珍珠素 (perlecan) 即屬於 HSPG 的一種

，本研究是在探討基底膜中硫酸肝素蛋白多醣 (heparan sulfate proteoglycan) 珍珠素 (perlecan) 在神經系統所扮演的角色與功能，先前的研究中證實重組的 人類珍珠素蛋白多醣第五片段 (human perlecan domain V; H.Pln DV) 在以 lami nin 為基材貼覆的環境下具有促進背根神經節神經纖維生長的能力，因此本實 驗主要目的是以重組基因技術 (recombinant DNA technology) 將 perlecan domai n V 分成兩個含類似組成的片段 (H.Pln DV-I, H.Pln DV-II) 探討其對神經纖維 生長之影響及作用機轉，研究結果顯示兩片段都會促進神經纖維的生長，先 前文獻中顯示 perlecan 會與乙醯膽鹼酯酵素 (AchE) 有交互作用且 AchE 有促 進神經纖維生長之作用，本研究第二部分探討 perlecan 與 AchE 的作用是否是 透過 perlecan domain V 。結果顯示 H.Pln DV, DV-I, DV-II 可直接與 AchE 結 合。推測 H.Pln DV 可能藉由與 AchE 的結合促進神經纖維的生長的。

探討人類珍珠素蛋白多醣第五片 段促進神經纖維生長之作用

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The major components of basement membrane (BM) are type IV collagen, laminin and proteoglycans. The basement membrane fuctions as a scaffold t o maintain the morphology of epithelium and endothelial cells. The major ty pe of proteoglycan in BM is heparan sulfate proteoglycan (HSPG). Previous study has shown that HSPG can promote the neurite outgrowth. Our studies are to understand the role of perlecan, which is the predominant HSPG in B M in the neurite outgrowth. Our previous study showed that recomibnant hu man perlecan domain V could promote the neurite outgrowth of dorsal root ganglion (DRG) neurons in laminin coating. Here we divided the perlecan d omain V (H.Pln DV) into two smaller fragment to identify the minimum fra gment required for promoting the neurite outgrowth. The result showed that both of these two smaller fragments derived from perlecan domain V could promote the neurite outgrowth of DRG neurons. Previously study showed th at perlecan and acetylcholinesterase (AchE) could interact with each other a nd AchE could promote the neurite outgrowth. Our further studies showed t hat H.Pln DV, H.Pln DV-I, H.Pln DV-II could bind to AchE. It suggests tha t H.Pln DV promoting the neurite outgrowth may be mediated by binding to AchE.

Studies of the effect of human perlecan domain V

on promoting neurite outgrowth