臺灣博碩士論文加值系統

血管再阻塞的病理機制主要歸因於血管平滑肌細胞的過度增殖和遷移。在以下的研究中，我們去探討甘露飲及桑黃萃取物hispolon對於抑制血管再阻塞的相關機轉。
在第一個章節中，主要是在探索頸動脈氣球擴張術大鼠模型中甘露飲對新生內膜形成及經由胎牛血清誘導的血管平滑肌細胞遷移的分子機制的。我們的研究結果指出甘露飲能顯著減少新生內膜的厚度。因此，本研究的目的是探討甘露飲的抗增生和抗轉移的效果，並探討其分子機制。在結果中，我們觀察到使用沒有細胞毒殺性的甘露飲有抑制血管平滑肌細胞的遷移能力。 並且能使分泌的基質金屬蛋白酶-2和基質金屬蛋白酶-9的表現量下降。而甘露飲對於血管平滑肌細胞遷移的抑制是通過其抑制磷酸化FAK、ERK1 / 2及PI3K/AKT的負調控作用。
因此我們的研究結果證實，甘露飲能抑制血管平滑肌細胞的轉移，並藉由MMP-2、MMP-9表現量降低去抑制FAK信號轉導通路及PI3K/AKT的路徑。經由這些數據，可推測甘露飲可能通過抑制血管平滑肌細胞遷移，而誘導細胞凋亡來抑制新生內膜的增生。
第二個部分是在探討hispolon對於大鼠頸動脈模式中抑制內膜增生及經由胎牛血清刺激誘導的平滑肌細胞的相關分子機制探討。經由實驗數據指出hispolon顯著抑制血管平滑肌細胞遷移，降低了基質金屬蛋白酶-2（MMP-2）和基質金屬蛋白酶9（MMP-9）的表達和分泌，並且增加了基質金屬蛋白酶組織抑制因子的表現。而在非細胞毒性劑量的hispolon作用下，其對血管平滑肌細胞遷移的抑製作用是通過其負調控作用磷酸化ERK1 / 2、PI3K/AKT。這些結果顯示，hispolon能夠降低MMP-9來抑制FAK信號轉導通路和PI3K/AKT的活性，進而抑制血管平滑肌細胞的遷移。此外，hispolon能夠通過抑制血管平滑肌細胞增殖和遷移，阻斷氣球擴張術後所誘導的新生內膜增生。

The pathological mechanism of restenosis is attributed primarily to excessive proliferation and migration of vascular smooth muscle cells. In these studies, we are trying to invesgate the anti-restenosis of Gan-Lu-Yin (GLY) and hispolon.
Chapter 1 was to explore the effects of GLY on neointimal formation in a rat model of carotid artery balloon injury and on the migration of vascular smooth muscle cells induced by fetal bovine serum and its molecular mechanisms. Our results showed that ethanol extract of GLY significantly decreased the thickness of neointimal, and the ratio of neointimal to media was also reduced. Therefore, the purpose of this study was to elucidate the antiproliferative and antimetastatic effects of GLY and to investigate its molecular mechanisms. In this study, we first observed that GLY exerted a dose-dependent inhibitory effect on migration (transwell assay) and motiliy (wound healing assay) of vascular smooth muscle cells in the absence of cytotoxicity. GLY decreased the expression and secretion of matrix metalloproteinase-2 (MMP-2) and MMP-9 in a concentration-dependent manner. The inhibition by non-cytoxic doses of GLY of vascular smooth muscle cells migration was mediated through its negative regulatory effects on phosphorylated extracellular signaling-regulating kinase1/2 (ERK1/2), phosphatidylinositol-3-kinase/serine/threonine protein kinase (PI3K/AKT), and phosphorylated focal adhesion kinase (FAK). Our results demonstrate that GLY can inhibit the metastasis of vascular smooth muscle cells by reduced expression of MMP-2 and MMP-9 through the suppression of the FAK signaling pathway and of the activity of PI3K/AKT. These data suggest that GLY might blocks injury-induced neointimal hyperplasia via the inhibition of vascular smooth muscle cells migration, without inducing apoptosis. These observations provide a mechanism of GLY in attenuating cell migration, thus as a potential intervention for restenosis.
And chapter 2 was to investigate the preventive effects of hispolon on balloon injury-induced neointimal formation and showed potent activity in inhibiting fetal bovine serum-induced vascular smooth muscle cells outgrowth. Hispolon significantly inhibited vascular smooth muscle cells migration as shown by trans-well assays. Hispolon decreased the expression and secretion of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9).The expression of the endogenous inhibitors of these proteins, namely, tissue inhibitors of MMP (TIMP-1 and TIMP-2), increased. The inhibition by non-cytoxic doses of hispoln of vascular smooth muscle cells migration was through its negative regulatory effects on phosphorylated FAK, phosphorylated ERK1/2, and PI3K/AKT. These results demonstrate that hispolon can inhibit the migration of vascular smooth muscle cells by reduced expression of MMP-9 through the suppression of the FAK signaling pathway and of the activity of PI3K/AKT. Our data suggest that hispolon might block balloon injury-induced neointimal hyperplasia via the inhibition of vascular smooth muscle cells proliferation and migration, without inducing apoptosis.

摘要 i
Abstract iii
Contents v
Figure contents vii
1.General Introduction 1
2. Materials and Methods 5
3. A chinese herbal formula “Gan-Lu-Yin” suppresses vascular smooth muscle cell migration by inhibiting matrix metalloproteinase-2/9 through the PI3K/AKT and ERK signaling pathways.
3.1. Brief introduction 13
3.2. Results 14
3.3. Discussion 17
3.4. Figures and figure legends 25
4. Hispolon attenuates balloon-injured neointimal formation and modulates vascular smooth muscle cell migration via AKT and ERK phosphorylation
4.1. Brief introduction 34
4.2. Results and Discussion 36
4.3. Figures and figure legends 44
5. Conclusion 57
6. References 58

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