鞘氨醇1-磷酸鹽對於建構人工血管影響之探討

Abstract

心血管疾病是由心臟及血管病變所引起，也是國內十大死因之一。目前治療方式主要為繞道移植手術。因此，移植材料的選擇是一個很重要的議題。其中廣泛被認知的人類臍靜脈由於是天然血管因而被認為適合用於移植。在本實驗中,我們利用SDS來清除臍靜脈上面原有的細胞群,並利用FBS進一步清除殘留的核酸以防止免疫反應的發生並藉由組織切片及DNA定量的方式來確認細胞清除的效果。繼而再將臍靜脈內皮細胞 (Human umbilical vein endothelial cells, HUVECs)及內皮前驅細胞 (Endothelial progenitor cells, EPCs)種回臍靜脈。除了讓細胞在靜止及動態的環境下貼附，我們另外在種細胞的實驗中加入鞘氨醇1-磷酸鹽 (Sphingosine1-phosphate, S1P)，S1P是一種水解磷酯脂，其會調節細胞的分化、遷移及貼附。由實驗結果顯示，無論是靜態或是動態的環境下，S1P會增強內皮細胞貼附於去細胞的臍靜脈。之後的凝血時間測試顯示了有種細胞的臍靜脈有較強的抗凝血能力。我們的實驗架構中顯示此實驗提供了不同的製作血管的方法並減少人工血管的製作所需的時間。

Cardiovascular disease (CVD), disorders of the heart and blood vessels, is the leading cause of mortality and morbidity. Arterial bypass graft is a primary therapy for the CVD patients. Therefore, choosing a proper material for arterial bypass grafting is a critical issue. Human umbilical vein (HUV) is a natural vessel and has been suggested as a suitable scaffold for vascular tissue engineering. In this study, we successfully de-cellularized HUV by using sodium dodecyl sulfate (SDS) and further incubated with medium contains 12% FBS to remove the residual nucleic acid. After the de-cellularization procedures, human umbilical vein endothelial cells (HUVECs) and endothelial progenitor cell (EPC) were then seeded back on de-cellularized HUV in the presence of 1uM sphingosine 1-phosphate (S1P) in static or rotational cultural conditions. S1P is a lysophospholipid (LPL) which binds to G protein-coupled receptors (GPCRs) and enhances endothelial cell proliferation, migration, and adhesion. Our results showed that S1P enhances endothelial attachment on de-cellularized HUV in static and rotational cultural conditions and reduces time required for preparation of vessel grafts. The finding potentially provides a novel strategy to develop vascular tissues.