動脈粥狀硬化為血管常見之疾病，其確切病理機制尚不明確，過去研究證據指出其原理為慢性發炎反應造成的血管病變的現象，且較常見於高血脂、高膽固醇、高血糖的患者。而其原因包含了因肥胖造成的脂肪組織堆積、毒素刺激如抽菸、酗酒及環境毒物皆會促使動脈粥狀硬化症的發生。臨床上已發展出許多手術用來治療這類疾病，像是氣球擴張術、動脈內膜切除或血管繞道手術。但約有四成的患者，在術後仍發生血管內膜增生，使血管再窄化。動脈粥狀硬化在目前治療環境下，仍缺乏一有效且能根治的方法。目前相關研究指出，靈芝具備的藥理特性可以運用在多項疾病當中，包括高血壓、高血脂、糖尿病和癌症等。我們利用小鼠頸動脈結紮手術，模擬低血流剪力與血流混亂導致內膜增生及動脈粥狀硬化症的發生，並且給予靈芝與靈芝三萜類觀察其對血管的保護作用。小鼠在執行頸動脈手術後一週，觀察到血管內膜產生增生現象；給予靈芝粉(300 mg/kg/day)之後內膜增生現象達到改善。我們又進一步將手術時間提升至兩週，並給予小鼠靈芝三萜類(300 mg/kg/day)，發現血管內膜增生現象在兩週後更加顯著，且在給與靈芝三萜類之後此現象得到明顯得到改善。為詳加瞭解靈芝三萜類作用機制，進一步利用組織染色觀察結果發現，在給與小鼠靈芝三萜類後，顯著降低血管內的氧化壓力；且靈芝三萜類可抑制內皮素(endotheln-1)與細胞黏附因子(VCAM-1)的表現，進而減緩內皮的細胞功能異常。內皮細胞在正常的情況下，可以降低血管內單核球(巨噬細胞)的浸潤而減緩血管病變後的發炎反應。此外，血管內高氧化壓力與浸潤的巨噬細胞所分泌的生長因子，皆會誘使平滑肌細胞在內膜的不正常增生，進而導致內膜增生。我們實驗指出，靈芝三萜類可以有效減少小鼠血管內膜增生的現象，且降低血管病變後產生的氧化壓力，同時保護內皮細胞，進而抑制巨噬細胞浸潤與平滑肌細胞在內膜層的過度增生。預期在未來，可以提升靈芝在動脈粥狀硬化這項疾病研究的重要性。

Neointima formation is common in various vascular diseases, such as atherosclerosis, and restenosis. Clinically, several types of surgery aim to enlarge narrowing vessels due to atherosclerosis, such as balloon angioplasty, endarterectomy and bypass grafting. However, over 40% of patients develop restenosis 6 months after surgery, which caused by the maladaptation of endothelial cells (ECs). EC dysfunction resulting from low shear stress or other risk factors, also it’s an initial progression of vascular diseases. EC dysfunction is associated with increased reactive oxygen species (ROS) levels, low-density lipoprotein (LDL) uptake, monocytes infiltration, and SMC proliferation. Ganoderma tsugae (GT) has been used for medicinal purposes in Asian countries for centuries. GT possesses various biological activities, including anti-oxidation, anti-inflammation and immunomodulation. However, the therapeutic potential of GT in intimal thickening is very limited. Triterpenoids of GT, composed of ganoderic acids/alcohols (GAs), are the bioactive ingredients for anti-hypertension, anti-oxidation and anti-inflammation. In this study, we intended to the therapeutic potential of GAs in neointima formation induced by common carotid artery ligation. This animal model creates disturbed flow and low shear stress in the arteries, and promotes neointima formation. Our result shows that the treatment of GT or GAs attenuates neointima formation induced by carotid artery ligation model. GAs treatment significantly suppressed the production of ROS, and prevented EC dysfunction by decreased the expression of vascular cell adhesion molecule 1 (VCAM-1) and endothelin 1 (ET-1). This result associated the infiltration of macrophages, and the proliferation of SMCs. Our findings suggests that GAs is the medicinal components of Lingzhi to prevent neointima formation.

中文摘要-----------------------------------------------------------------------------------------------------------i
Extended Abstract-----------------------------------------------------------------------------------------------ii
致謝-----------------------------------------------------------------------------------------------------------------v
內文目錄----------------------------------------------------------------------------------------------------------vi
圖表目錄--------------------------------------------------------------------------------------------------------viii
縮寫表-------------------------------------------------------------------------------------------------------------ix
第一章 前言------------------------------------------------------------------------------------------------------1
第一節 動脈粥狀硬化 Atherosclerosis-------------------------------------------------------------1
一、 血管結構組成---------------------------------------------------------------------------------1
二、 動脈粥狀硬化之成因------------------------------------------------------------------------1
三、 動脈粥狀硬化與血流紊亂-----------------------------------------------------------------3
四、 動脈粥狀硬化的治療-----------------------------------------------------------------------4
第二節 頸動脈結紮手術 Carotid Artery Ligation-----------------------------------------------5
第三節 靈芝(Ganoderma)與三萜類(Triterpenoids)----------------------------------------------5
研究動機-----------------------------------------------------------------------------------------------------------8
第二章 材料與方法--------------------------------------------------------------------------------------------9
第一節 材料-----------------------------------------------------------------------------------------------9
一、 藥品----------------------------------------------------------------------------------------------9
二、 藥品配置--------------------------------------------------------------------------------------11
三、 實驗材料--------------------------------------------------------------------------------------13
四、 儀器設備--------------------------------------------------------------------------------------15
第二節 方法---------------------------------------------------------------------------------------------16
一、 實驗動物--------------------------------------------------------------------------------------16
二、 組織製備與切片----------------------------------------------------------------------------18
三、 組織染色--------------------------------------------------------------------------------------19
四、 定量分析--------------------------------------------------------------------------------------23
第三章 結果----------------------------------------------------------------------------------------------------24
第一節 靈芝對於小鼠頸動脈結紮術後的影響------------------------------------------------24
靈芝保護血管免於頸動脈結紮手術所造成的血管內膜新生-------------------24
第二節 靈芝三萜類對於小鼠頸動脈結紮術後的影響--------------------------------------25
一、 靈芝三萜類保護血管免於頸動脈結紮手術所造成的血管內膜新生---------------------------------------------------------------------------------------------------25
二、 靈芝三萜類保護血管免於受到氧化壓力--------------------------------------------26
三、 靈芝三萜類抑制內皮細胞損傷---------------------------------------------------------27
四、 靈芝三萜類減緩血管發炎反應---------------------------------------------------------28
五、 靈芝三萜類減少平滑肌細胞在內膜層的過度增生--------------------------------29
第四章 討論----------------------------------------------------------------------------------------------------30
第五章 結論----------------------------------------------------------------------------------------------------36
第六章 參考文獻----------------------------------------------------------------------------------------------37
第七章 圖表----------------------------------------------------------------------------------------------------42

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