七層塔萃取物抗氧化及減低粥狀動脈硬化發展之研究

隨著科技時代的來臨，人們飲食習慣也隨之改變，享受其帶來的便利卻也伴隨著許多文明病的發生。過量膽固醇的攝取導致低密度脂蛋白 (low-density lipoprotein；LDL) 囤積在動脈血管壁上，經氧化形成氧化型低密度脂蛋白 (oxidative modification of low-density lipoprotein；oxLDL)進而引發血管內皮細胞受損。過去文獻也曾指出粥狀動脈硬化的形成與氧化型低密度脂蛋白的累積有著密切的關係。草本植物七層塔 (Ocimum gratissimum) 有抑菌、抗發炎和抗氧化的功效，對於能否有效抑制 LDL氧化並抑制粥狀動脈硬化形成及其詳細的作用機制到目前為止還不清楚。在本篇論文中我們首先要探討七層塔是否可以保護低密度脂蛋白不受自由基的傷害，減少氧化型低密度脂蛋白的產生，降低誘發巨噬細胞吞噬氧化型低密度脂蛋白所產生的泡沫細胞，並且保護氧化型低密度脂蛋白對人類臍帶靜脈內皮細胞 (HUVECs) 所造成的病變。在抑制 LDL 氧化實驗中，透過 relative electrophoretic mobility (REM) shift assays、electrophoresis of ApoB fragmentation、lipid peroxidation assay、證明七層塔能有效抑制硫酸銅所誘導 LDL 的氧化。又利用 (JC-1 stain) 螢光陽離子結合粒線體膜電位檢測以及細胞凋亡檢測 (DAPI stain) 觀察細胞型態的表現，也證實在七層塔的作用下氧化型低密度脂蛋白對 HUVECs 所造成的傷害能有效被抑制，而在西方墨點 (Western blot) 的相關蛋白分析中也表現出一致的現象，所以歸納出七層塔能有效抑制低密度脂蛋白的氧化和減少巨噬細胞吞噬氧化型低密度脂蛋白所產生的泡沫細胞，進而達到保護人類血管內皮細胞。

Today, people enjoy the convenience it brings but also accompanied by the occurrence of many diseases of civilization. Too much cholesterol lead to low-density lipoprotein accumulation in the arterial wall, damage caused by vascular endothelial formation of oxidized low-density lipoprotein. The formation of atherosclerosis correlate with oxidized low-density lipoprotein accumulation. The Ocimum gratissimum extract (OGE) have antibacterial, anti-inflammatory and anti-oxidant effect, for the development of atherosclerosis and its detailed mechanism of action is unclear so far whether inhibition of atherosclerosis. In this thesis, we shows, the OGE can protect against free radical damage of low-density lipoprotein, reduce the production of oxidized low-density lipoprotein, the foam cells and macrophages induced by oxidized low-density lipoprotein, and to protect the oxidized low-density lipoprotein lesions caused by human vascular endothelial cells. In the inhibition of LDL oxidation experiments,through the relative electrophoretic mobility (REM) shift assays, electrophoresis of ApoB fragmentation, lipid peroxidation assay, prove that the OGE can effectively inhibit the copper sulfate-induced LDL oxidation and (JC-1 stain) fluorescent cation binding mitochondrial membrane potential detection and apoptosis. Death detection (DAPI stain), the cells were observed patterns of performance, also confirmed that the damage caused by HUVECs reply changed in the OGE under the action of oxidized low-density lipoprotein, and in the Western blot the experiment also showed a consistent phenomenon. In conclusion, The OGE can effectively inhibit the oxidation of low-density lipoprotein and reduce macrophage phagocytosis of oxidized low-density lipoprotein foam cells, and thus achieve the protection of human vascular endothelial cells.