臺灣博碩士論文加值系統

指導教授推薦書
口試委員審定書
誌謝…………………………………………………………………… iii
中文摘要……………………………………………………………………V
英文摘要…………………………………………………………………vi
目錄………………………………………………………………………vii
圖表目錄……………………………………………………………………x
縮寫表……………………………………………………………………xi
一、前言……………………………………………………………………1
1.1 研究背景 …………………………………………………………1
1.1.1動脈粥狀硬化(atherosclerosis)和血管再狹窄(restenosis)………1
1.1.2bFGF對血管平滑肌細胞增生和遷移的調控……………………3
1.1.3漢黃芩素的藥理作用………………………………………………8
1.2 研究目的…………………………………………………12
二、材料與方法 …………………………………………………………13
2.1 材料………………………………………………………………13
2.1.1實驗抗體材料………………………………………………………13
2.2 實驗方法………………………………………………………13
2.2.1細胞培養……………………………………………………………13
2.2.2蛋白質定量…………………………………………………………14
2.2.3西方點墨法…………………………………………………………15
2.2.4細胞計數……………………………………………………………15
2.2.5Zymography ………………………………………………………16
2.2.6細胞移行分析………………………………………………………16
2.2.7數據的分析方法……………………………………………………17
三、實驗結果………………………………………………………………18
3.1.1漢黃芩素抑制bFGF誘導血管平滑肌細胞的增生………………18
3.1.2漢黃芩素和訊號抑制劑對bFGF誘導血管平滑肌增生的影響…19
3.2.1漢黃芩素抑制bFGF誘導血管平滑肌細胞遷移…………………19
3.2.2漢黃芩素和訊號抑制劑抑制bFGF誘導血管平滑肌遷移的影響20
3.3 漢黃芩素抑制bFGF誘導的ERK1/2磷酸化……………………20
3.4 漢黃芩素抑制bFGF誘導活化轉錄因子Egr-1的活性…………21
3.5 漢黃芩素抑制bFGF誘導細胞週期素D1和A蛋白量………… 22
3.6 漢黃芩素對bFGF誘導P21、P27的影響…………………………23
3.7 漢黃芩素對bFGF誘導MMP-2/MMP-9活性的影響……………23
3.8 漢黃芩素可能作用模式………………………………………24
四、討論……………………………………………………………………35
五、結論……………………………………………………………………40
參考文獻…………………………………………………………………41
圖表目錄
圖3.1.1漢黃芩素對bFGF誘導血管平滑肌細胞增生的影響…………25
圖3.1.2漢黃芩素和各種抑制劑對bFGF誘導血管平滑肌細胞增生…26
圖3.2.1漢黃芩素對bFGF誘導血管平滑肌細胞遷移的影響…………27
圖3.2.2漢黃芩素和各種抑制劑對bFGF誘導血管平滑肌遷移的影響28
圖3.3 漢黃芩素對bFGF誘導血管平滑肌細胞ERK磷酸化的影響29
圖3.4.1漢黃芩素對bFGF誘導活化轉錄因子Egr-1的影響…………30
圖3.4.2漢黃芩素對bFGF誘導活化轉錄因子NF-κB的影響………31
圖3.5 漢黃芩素對bFGF誘導血管平滑肌細胞週期蛋白的影響……32
圖3.7 漢黃芩素抑制bFGF誘導MMP-2/MMP-9影響………………33
圖3.8 漢黃芩素可能作用模式………………………………………34

Abukhdeir AM, Park BH. (2008) P21 and p27: roles in carcinogenesis and drug resistance. Expert Rev Mol Med. 10:e19.
Basilico C, Moscatelli D. (1992) The FGF family of growth factors and oncogenes. Adv Cancer Res. 59:115-165.
Baird A. (1994) Fibroblast growth factors: activities and significance of non-
neurotrophin neurotrophic growth factors. Curr Opin Neurobiol. 4:78-86.
Cakir Y, Ballinger SW. (2005) Reactive species-mediated regulation of cell signaling and the cell cycle: the role of MAPK. Antioxid Redox Signal. 7:726-740.
Chang YL, Shen JJ, Wung BS, Cheng JJ, Wang DL. (2001) Chinese herbal remedy wogonin inhibits monocyte chemotactic protein-1 gene expression in human endothelial cells. Mol Pharmacol. 60:507-513.
Chen LG, Hung LY, Tsai KW, Pan YS, Tsai YD, Li YZ, Liu YW. (2008) Wogonin, a bioactive flavonoid in herbal tea, inhibits inflammatory cyclooxygenase-2 gene expression in human lung epithelial cancer cells.Mol Nutr Food Res. 52:1349-1357.
Chi YS, Lim H, Park H, Kim HP. (2003) Effects of wogonin, a plant flavone from Scutellaria radix, on skin inflammation: in vivo regulation of inflammation-associated gene expression. Biochem Pharmacol. 66:1271-78.
Cho J, Lee HK. (2004) Wogonin inhibits excitotoxic and oxidative neuronal damage in primary cultured rat cortical cells. Eur J Pharmacol. 485:105-110.
Dong LH, Wen JK, Miao SB, Jia Z, Hu HJ, Sun RH, Wu Y, Han M. (2010) Baicalin inhibits PDGF-BB-stimulated vascular smooth muscle cell proliferation through suppressing PDGFRβ-ERK signaling and increase in p27 accumulation and prevents injury-induced neointimal hyperplasia. Cell Res. 20:1252-1262.
Fallon JF, López A, Ros MA, Savage MP, Olwin BB, Simandl BK. (1994) FGF-2: apical ectodermal ridge growth signal for chick limb development. Science. 264:104-107.
Fischman DL, Leon MB, Baim DS, Schatz RA, Savage MP, Penn I, Detre K, Veltri L, Ricci D, Nobuyoshi M. (1994) A randomized comparison of coronary-stent placement and balloon angioplasty in the treatment of coronary artery disease. Stent Restenosis Study Investigators. N Engl J Med. 331:496-501.
Galis ZS, Khatri JJ. (2002) Matrix metalloproteinases in vascular remodeling and atherogenesis: the good, the bad, and the ugly. Circ Res. 90:251-262.
Gao Z, Huang K, Yang X, Xu H. (1999) Free radical scavenging and antioxidant activities of flavonoids extracted from the radix of Scutellaria baicalensis Georgi. Biochimica et Biophysica Acta. 1472:643-650.
Gao Z, Huang K, Xu H. (2001) Protective effects of flavonoids in the roots of Scutellaria baicalensis Georgi against hydrogen peroxide-induced oxidative stress in HS-SY5Y cells. Pharmacol Res. 43:173-178.
Gospodarowicz D. (1975) Purification of a fibroblast growth factor from bovine pituitary. J Biol Chem. 250:2515-2520.
Grube E, Silber S, Hauptmann KE, Mueller R, Buellesfeld L, Gerckens U, Russell ME. (2003) TAXUS I: six- and twelve-month results from a randomized, double-blind trial on a slow-release paclitaxel-eluting stent for de novo coronary lesions. Circulation. 107:38-42.
Guillemot L, Levy A, Raymondjean M, Rothhut B. (2001) Angiotensin II-induced transcriptional activation of the cyclin D1 gene is mediated by Egr-1 in CHO-AT(1A) cells. J. Biol. Chem. 276:39394-39403.
Guo Q, Zhao L, You Q, Yang Y, Gu H, Song G, Lu N, Xin J. (2007) Anti-hepatitis B virus activity of wogonin in vitro and in vivo. Antiviral Res. 74:16-24.
Han W, Liu GN. (2010) EGR-1 decoy ODNs inhibit vascular smooth muscle cell proliferation and neointimal hyperplasia of balloon-injured arteries in rat. Life Sci. 86:234-243.
Huang RL, Chen CC, Huang HL, Chang CG, Chen CF, Chang C, Hsieh MT.
(2000) Anti-hepatitis B virus effects of wogonin isolated from Scutellaria baicalensis. Planta Med. 66:694-698.
Imamura M, Kanematsu A, Yamamoto S, Kimura Y, Kanatani I, Ito N, Tabata Y, Ogawa O. (2007) Basic fibroblast growth factor modulates proliferation and collagen expression in urinary bladder smooth muscle cells. Am J Physiol Renal Physiol. 293:F1007-1017.
Khachigian LM. (2006) Early growth response-1 in cardiovascular pathobiology. Circ. Res. 98:186-191.
Kimura Y, Okuda H, Ogita Z. (1997) Effects of flavonoids isolated from scutellariae radix on fibrinolytic system induced by trypsin in human umbilical vein endothelial cells. J Nat Prod. 60:598-601.
Kim EJ, Kang D, Han J. (2011) Baicalein and wogonin are activators of rat TREK-2 two-pore domain K+ channel. Acta Physiol (Oxf). 202:185-192.
Kitamura K, Honda M, Yoshizaki H, Yamamoto S, Nakane H, Fukushima M, Ono K, Tokunaga T. (1998) Baicalin, an inhibitor of HIV-1 production in vitro. Antiviral Research. 37:131-140.
Lam TL, Lam ML, Au TK, Ip DT, Ng TB, Fong WP, Wan DC. (2000) A comparison of human immunodeficiency virus type-1 protease inhibition activities by the aqueous and methanol extracts of Chinese medicinal herbs. Life Sci. 67:2889-2896.
Lee H, Kim YO, Kim H, Kim SY, Noh HS, Kang SS, Cho GJ, Choi WS, Suk K. (2003) Flavonoid wogonin from medicinal herb is neuroprotective by inhibiting inflammatory activation of microglia. The FASEB Journal. 13:1943-1964.
Lee SO, Jeong YJ, Yu MH, Lee JW, Hwangbo MH, Kim CH, Lee IS. (2006) Wogonin suppresses TNF-a-induced MMP-9 expression by blocking the NF-jB activation via MAPK signaling pathways in human aortic smooth muscle cells. Biochemical and Biophysical Research Communications. 351:118-125.
Li BQ, Fu T, Gong WH, Dunlop N, Kung H, Yan Y, Kang J, Wang JM. (2000) The flavonoid baicalin exhibits anti-inflammatory activity by binding to chemokines. Immunopharmacology. 49:295-306.
Lille S, Daum G, Clowes MM, Clowes AW. (1997) The regulation of p42/p44 mitogen-activated protein kinases in the injured rat carotid artery. J Surg Res. 70:178-186.
Lin CM, Chen YH, Ong JR, Ma HP, Shyu KG, Bai KJ. (2012) Functional role of wogonin in anti-angiogenesis. Am J Chin Med. 40:415-427.
Liu B, Fisher M, Groves P. (2002) Down-regulation of the ERK1 and ERK2 mitogen-activated protein kinases using antisense oligonucleotides inhibits intimal hyperplasia in a porcine model of coronary balloon angioplasty. Cardiovasc Res.54:640-648.
Liu JJ., Huang TS., Cheng WF., Lu FJ. (2003) Baicalein and baicalin are potent inhibitors of angiogenesis: Inhibition of endothelial cell proliferation, migration and differentiation. Int J Cancer. 106:559-565.
Logan A, Frautschy SA, Baird A. (1991) Basic fibroblast growth factor and central nervous system injury. Ann N Y Acad Sci. 638:474-476.
Ma SC, Du J, But PP, Deng XL, Zhang YW, Ooi VE, Xu HX, Lee SH, Lee SF. (2002) Antiviral Chinese medicinal herbs against respiratory syncytial virus. J Ethnopharmacol. 79:205-211.
McAvoy JW, Chamberlain CG, de Iongh RU, Richardson NA, Lovicu FJ.
(1991) The role of fibroblast growth factor in eye lens development. Ann N Y Acad Sci. 638:256-274.
Nagai T, Suzuki Y, Tomimori T, Yamada H. (1995) Antiviral activity of plant flavonoid, 5,7,4'-trihydroxy-8-methoxyflavone, from the roots of Scutellaria baicalensis against influenza A (H3N2) and B viruses. Biol Pharm Bull. 18:295-299.
Ornitz DM, Xu J, Colvin JS, McEwen DG, MacArthur CA, Coulier F, Gao G, Goldfarb M. (1996) Receptor specificity of the fibroblast growth factor family. J Biol Chem. 271:15292-15297.
Park BK, Heo MY, Park H, Kim HP. (2001) Inhibition of TPA-induced cyclooxygenase-2 expression and skin inflammation in mice by wogonin, a plant flavone from Scutellaria radix. Eur J Pharmacol. 425:153-157.
Popma JJ, Topol EJ. (1990) Factors influencing restenosis after coronary angioplasty. Am J Med. 88:16N-24N.
Rider V, Piva M, Cohen ME, Carlone DL. (1995) Alternative splicing and differential targeting of fibroblast growth factor receptor 1 in the pregnant rat uterus.Endocrinology. 136:3137-3145.
Riley BB, Savage MP, Simandl BK, Olwin BB, Fallon JF. (1993) Retroviral expression of FGF-2 (bFGF) affects patterning in chick limb bud. Development.118:95-104.
Ross R. (1993) The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature. 362:801-809.
Ross R. (1999) Atherosclerosis--an inflammatory disease. N Engl J Med. 340:115-126.
Santiago FS, Lowe HC, Day FL, Chesterman CN., Khachigian LM. (1999) Early growth response factor-1 induction by injury is triggered by release and paracrine activation by fibroblast growth factor-2. Am J Pathol.154:937-944.
Sapienza P, di Marzo L, Borrelli V, Sterpetti AV, Mingoli A, Piagnerelli R, Cavallaro A. (2004) Basic fibroblast growth factor mediates carotid plaque instability through metalloproteinase-2 and -9 expression. Eur J Vasc Endovasc Surg. 28:89-97.
Schwartz SM, Liaw L. (1993) Growth control and morphogenesis in the development and pathology of arteries. J Cardiovasc Pharmacol.21:S31-49.
Schröder K, Helmcke I, Palfi K, Krause KH, Busse R, Brandes RP. (2007) Nox1 mediates basic fibroblast growth factor-induced migration of vascular smooth muscle cells. Arterioscler Thromb Vasc Biol. 27:1736-1743.
Serruys PW, de Jaegere P, Kiemeneij F, Macaya C, Rutsch W, Heyndrickx G, Emanuelsson H, Marco J, Legrand V, Materne P. (1994) A comparison of balloon-expandable-stent implantation with balloon angioplasty in patients with coronary artery disease. Benestent Study Group. N Engl J Med. 331:489-495.
Serruys PW, Regar E, Carter AJ. (2002) Rapamycin eluting stent: the onset of a new era in interventional cardiology. Heart. 87:305-307.
Skaletz-Rorowski A, Waltenberger J, Müller JG, Pawlus E, Pinkernell K, Breithardt G. (1999) Protein kinase C mediates basic fibroblast growth factor
-induced proliferation through mitogen-activated protein kinase in coronary smooth muscle cells. Arterioscler Thromb Vasc Biol.19:1608-1614
Skaletz-Rorowski A, Eschert H, Leng J, Stallmeyer B, Sindermann JR, Pulawski E, Breithardt G. (2005) PKC delta-induced activation of MAPK pathway is required for bFGF-stimulated proliferation of coronary smooth muscle cells. Cardiovasc Res. 67:142-150.
Unsicker K, Engels S, Hamm C, Lüdecke G, Meier C, Renzing J, Terbrack HG, Flanders K. (1992) Molecular control of neural plasticity by the multifunctional growth factor families of the FGFs and TGF-beta s. Ann Anat. 174:405-407.
Wang F, Song X, Zhou M, Wei L, Dai Q, Li Z, Lu N, Guo Q. (2013) Wogonin inhibits H2O2-induced vascular permeability through suppressing the phosphorylation of caveolin-1. Toxicology. 305:10-19.
Weng CJ, Chau CF, Hsieh YS, Yang SF, Yen GC. (2008) Lucidenic acid inhibits PMA-induced invasion of human hepatoma cells through inactivating MAPK/ERK signal transduction pathway and reducing binding activities of NF-kappaB and AP-1. Carcinogenesis. 29:147-156.
Wu JA, Attele AS, Zhang L, Yuan CS. (2001) Anti-HIV activity of medicinal
herbs:usage and potential development. Am J Chin Med. 29:69-81.