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研究生:鍾幸君
研究生(外文):CHUNG HSING-CHUN
論文名稱:樟芝對平滑肌細胞的遷移及老鼠頸動脈新生內膜形成的抑制作用
論文名稱(外文):Novel inhibitory effect of Antrodia camphorata on smooth muscle cellmigration and carotid neointima formation in mice
指導教授:陳健祺
指導教授(外文):CHEN JIAN-CHYI
學位類別:碩士
校院名稱:南台科技大學
系所名稱:生物科技系
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:75
中文關鍵詞:樟芝大鼠動脈平滑肌細胞血小板衍生性生長因子遷移中層新生內膜
外文關鍵詞:Antrodia camphoratarat aortic smooth muscle cellplatelet-derived growth factormigrationmedianeointima.
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動脈粥狀硬化(Atherosclerosis)是心血管疾病中主要的疾病,在動脈粥狀病變與血管再窄化發展病程中,血管平滑肌細胞從中膜層遷移至新生血管內膜是血管形成病變的重要的因素。在血管受傷病變過程中,平滑肌細胞會由收縮態轉變為合成態,並由血管中層往內膜層增生和遷移,導致血管內膜增厚。樟芝(Antrodia camphorata,AC)是一種中國傳統常見的藥品,它具有抑制腫瘤細胞增生及遷移的作用。此論文欲研究樟芝對大鼠動脈平滑肌細胞 (Rat aortic smooth muscle cell;RASMC:A7r5)的增生及遷移是否也具有抑制效果;並且評估其對老鼠頸動脈受傷後之新生內膜生成的影響。MTT和BrdU 增生實驗主要是研究樟芝對平滑肌細胞增生作用的影響,由MTT和BrdU 實驗結果皆顯示樟芝可抑制血小板衍生生長因子(PDGF)所引起平滑肌細胞的增生作用。在Transwell 遷移實驗中,大鼠動脈平滑肌細胞(RASMC)先用樟芝萃取液或食鹽水處理後,然後血小板衍生生長因子(PDGF)加在Transwell的下層刺激細胞遷移,計算由上層遷移到下層的細胞數目。在wound scratch assay中,大鼠動脈平滑肌細胞(RASMC)用樟芝萃取液或食鹽水預先處理,使用pipette tips 刮除細胞;然後用血小板衍生生長因子(PDGF)刺激並培養24小時。此2個實驗結果皆顯示樟芝可顯著抑制血小板衍生生長因子(PDGF)所引起的平滑肌細胞遷移作用。動物實驗方面,將C57BL/6老鼠的左邊頸動脈在靠近分支處結紮手術,術後的老鼠用水及樟芝萃取液連續灌食4星期,新生內膜生成的嚴重性則用新生內膜/中層(neointima / media,N/M )比例來表示。在4星期後,以樟芝萃取液治療,經頸動脈結紮的老鼠其新生內膜生成較少(N/M ratio,水控制組, n=13 vs 樟芝 250 mg/kg, n=12 vs 樟芝1250 mg/kg, n=12;1.33±0.87 vs 0.83±0.45vs 0.63±0.32,P < 0.05)。實驗結果顯示樟芝對於血小板衍生生長因子(PDGF)引起平滑肌細胞的增生和遷移作用是個有效的抑制劑,另外以樟芝治療也可降低老鼠頸動脈結紮手術後的新生內膜的生成。
Atherosclerosis in western countries and Taiwan is the major cardiovascular diseases. The migration of vascular smooth muscle cells (VSMCs) from the media into the neointima is a key event in the development of atherosclerotic lesion and in restenosis. An important hallmark of vascular injury is the increased proliferation and migration of vascular smooth muscle cells (VSMCs) associated with the transformation from contractile to synthetic phenotype. Antrodia camphorate (AC) is a well-known traditional Chinese medicine. It has been shown to inhibit the proliferation and migration of cancer cells. This study examined whether AC could inhibit rat aortic smooth muscle cell (RASMC:A7r5) proliferation and migration and evaluated its effect on neointima formation in mouse carotid artery after injury. MTT and BrdU proliferation assay were used to evaluate AC effect on SMC proliferation. Both assays showed AC inhibits PDGF-induced SMC proliferation. In Transwell migration assay, RASMCs were treated with AC or saline. PDGF (10 ng/ml) was added to the lower compartment. Migrated cells on the lower side were fixed and counted. In wound scratch assay, RASMCs were pretreated with AC or saline and were seeded into confluent cell monolayers. They were wounded with pipette tips and stimulated with 10 ng/ml PDGF for 24 hrs. Both assays also showed AC could significantly inhibit PDGF-induced SMC migration. The left common carotid arteries of C57BL/6 mices were ligated near the carotid bifurcation. The mice were fed with water or AC for 4 wks. The severity of neointima was expressed as neointima/media (N/M) ratio. The AC-treated mice had less neointima formation at 4 wks after carotid ligation (N/M ratio, water, n=13 vs 250 mg/kg AC, n=12 vs 1250 mg/kg AC, n=12;1.33±0.87 vs 0.83±0.45 vs 0.63±0.32, P < 0.05). The data indicate that AC is an effective inhibitor of PDGF-induced RASMC proliferation and migration. AC treatment reduced neointima formation in mouse carotid ligation model.
中文摘要---------------------------------------------------------------------------------------- 1
英文摘要---------------------------------------------------------------------------------------- 2
誌謝---------------------------------------------------------------------------------------------- 4
目錄---------------------------------------------------------------------------------------------- 5
圖目錄------------------------------------------------------------------------------------------- 7
表目錄--------------------------------------------------------------------------------------------9
附錄-----------------------------------------------------------------------------------------------9
縮寫表------------------------------------------------------------------------------------------- 10
第一章 緒論------------------------------------------------------------------------------------ 11
第二章 研究動機與目的--------------------------------------------------------------------- 24
第三章 材料與方法--------------------------------------------------------------------------- 26
3-1 實驗材料----------------------------------------------------------------------------------- 27
3-2 實驗藥品----------------------------------------------------------------------------------- 27
3-3 實驗儀器----------------------------------------------------------------------------------- 29
3-4 實驗方法----------------------------------------------------------------------------------- 30
3-4-1 細胞存活率測定(MTT assay)------------------------------------------------------- 31
3-4-2 細胞存活率測定(BrdU assay) ----------------------------------------------------- 32
3-4-3 細胞遷移實驗(cell migration assay) -------------------------------------------- 32
3-4-4 細胞刮傷試驗(Wound scratching assay) -------------------------------------- 33
3-4-5 Gelatin Zymography----------------------------------------------------------------- 33
3-4-6 動物實驗(Animal test)--------------------------------------------------------------- 37
3-4-7 組織切片及染色----------------------------------------------------------------------- 38
3-4-8 血管檢查評估-------------------------------------------------------------------------- 39
3-4-9 統計分析法----------------------------------------------------------------------------- 40
第四章 實驗結果------------------------------------------------------------------------------ 41
第五章 討論------------------------------------------------------------------------------------ 44
第六章 參考文獻------------------------------------------------------------------------------ 47
圖表---------------------------------------------------------------------------------------------- 51
表格---------------------------------------------------------------------------------------------- 68
附錄---------------------------------------------------------------------------------------------- 69
自述---------------------------------------------------------------------------------------------- 75
王伯徹.具開發潛力食藥用菇介紹.食品工業.2000;32(5):1-17。

王伯徹.食藥用菇保健食品之開發.食品工業.2002;32(5):18-27。

王伯徹和陳啟楨.常見食藥用菇類介紹,食品工業發展研究所.1994。

陳勁初和呂鋒洲.靈芝之王:台灣樟芝,元氣齋出版社.2001。

張曉莉、薛文明、李恩慈、張瓊妃.樟芝子實體、菌絲體及相關產品三帖類差異
之探討.高苑技術學院-化學工程系.2002。

潘振澤.科學論文寫作與發表.藝軒圖書出版社.1994。

Bendeck MP, Zempo N, Clowes AW, Galardy RE, Reidy MA. Smooth muscle cell migration and matrix metalloproteinase expression after arterial injury in the rat. Circulation research. 1994;75:539–545.

Bonetti PO, Lerman LO, Lerman A. Endothelial dysfunction: a marker of Athero-sclerotic risk. Arteriosclerosis, thrombosis, and vascular biology. 2003;23(2):168-175.

Cheng JJ, Huang NK, Chang TT, Wang DL, LuMK. Study for anti-angiogenic activities of polysaccharides isolated from Antrodia cinnamomea in endothelial cell. Life Sciences. 2005;(76):3029–3042.

Hsiao G, Shen MY, Lin KH, Lan MH, Wu LY, Cho DS, Lin CH, Su CH, Sheu JR. Antioxidative and Hepatoprotective Effects of Antrodia camphorata Extract. Journal Agricultural and Food chemistry.2003;51:3302-3308.

Hseu YC, Chang WC, Hseu YT. Protection of oxidative damage by aqueous extract from Antrodia camphorata mycelia in normal human erythrocytes. Life Sciences. 2002;71:469-82.

Kumar A, Lindner V. Remodeling with neointima formation in the mouse carotid artery after cessation of blood flow. Arteriosclerosis, thrombosis,and vascular biology. 1997;17(10):2238-2244.

Laszik ZG, Zhou XJ, Ferrell GL, Silva FG, Esmon CT. Down-regulation of endothelial expression of endothelial cell protein C receptor and thrombomodulin in coronary atherosclerosis. 2001:797-802.

Lee IH, Huang RL, Chen CT, Chen HC, Hsu WC, Lu MK. Antrodia camphorata polysaccharides exhibit anti-hepatitis B virus effects. FEMS Microbiology Letters. 2002;209:63-67.

Li YH, Liu SL, Shi GY, Tseng GH, Liu PY, Wu HL. Thrombomodulin plays an important role in arterial remodeling and neointima formation in mouse carotid ligation model. Thromb Haemost. 2006;95:128-133.

Lin WC, Kuo SC, Lin WL, Fang HL, Wang BC. Filtrate of fermented mycelia from Antrodia camphorate reduces liver fibrosis induced by carbon tetrachloride in rats. World Journal of Gastroenterol. 2006;12(15):2369-2374.

Liu JJ, Huang TS, Hsu ML, et al. Antitumor effects of the partially purified Poly-Saccharides from Antrodia camphorata and the mechanism of its action. Toxicology and Applied Pharmacology. 2004;201:186-193.

Ma H, Calderon TM, Kessel T, Ashton AW, Berman JW. Mechanisms of hepatocyte growth factor-mediated vascular smooth muscle cell migration. Circulation research. 2003; 28:1066-1073.

Morigiwa,A.,Kitabatake,K.,Fujimoto,Y.,and Ikekawa,N.Angiotensin converting enzyme-inhibitory triterpenes from Ganoderma Iucidum.Chemical and Pharmaceutical Bulletin. 1986;34:3025-3028.

Nagase H, Woessner JF. Matrix metalloproteinases. Journal Biology Chemical. 1999;274:21491–21494.

Peng CC,Chen KC,Peng RY,Su CH,Hsieh-Li HM.Human urinary bladder cancer T24 cells are susceptible to the Antrodia camphorate extracts. Cancer Letters. 2006;243:109-119.

Peng CC, Chen KC, Peng RY, Chyau CC, Su CH, Hsieh-Li HM. Antrodia camphorate extract induces replicative senescence in superficial TCC, and inhibits the absolute migration capability in invasive bladder carcinoma cells. Journal Ethnopharmacol. 2007;109:93-103.

Popp O, Heidinger M, Ruiz-Heinrich L, Ries C, Jochum M, Gil-Parrado S. The calpastatin-derived calpain inhibitor CP1B reduces mRNA expression of matrix metalloproteinase-2 and -9 and invasion by leukemic THP-1 cells. Biology Chemical. 2003;384:951-958.

Ross R. Atherosclerosis an inflammatory disease. The New England journal of medicine. 1999;340(2):115-126.

Schwartz SM, Campbell GR, Campbell JH. Replication of smooth muscle cells in vascular disease. Circulation research. 1986;58(4):427-444.

Shapiro SD. Matrix metalloproteinases degradation of extracellular matrix:biological consequences. Current Opinine Cell Biology. 1998;10:602–608.

Shen YC, Wang YH, Chou YC, et al. Evaluation of the anti-inflammatory activity of zhankuic acids isolated from the fruiting bodies of Antrodia camphorata. Planta Medica. 2004;70:310-314.

Sliva D. Ganoderma lucidum in cancer research. Leukemia Research. 2006;30:767-768.

Song TY, Hsu SY, Yen GC. Induction of apoptosis in human hepatoma cells by mycelia of Antrodia camphorata in submerged culture. Journal of Ethnopharmacology. 2005;100:158–167.

Thyberg J, Blomgren K, Hedin U, Dryjski M. Phenotypic modulation of smooth muscle cells during the formation of neointimal thickenings in the rat carotid artery after balloon injury: an electron-microscopic and stereological study. Cell and tissue research.1995;281(3):421-433.

Wu SH, Ryvarden L, Chan TT. Antrodia camphorata (niu-chang-chih), new combination of a medicinal fungus in Taiwan. Bot Bull Acad Sin. 1997;38:273-275.

Wu H, Pan CL, Yao YC, Chang SS, Li SL, Wu TF. Proteomic analysis of the effect of Antrodia camphorata extract on human lung cancer A549 cell. Proteomics. 2006;6:826-835.

Yang HL, Chen CS, Chang WH, Lu FJ, Lai YC, Chen CC, Hseu TH, Kuo CT, Hseu YC. Growth inhibition and induction of apoptosis in MCF-7 breast cancer cells by Antrodia camphorate. Cancer Letters. 2006;231:215–227.

Yang HL, Hseu YC, Chen JY, et al. Antrodia camphorata in submerged culture protects low density lipoproteins against oxidative modification. The American journal of Chinese medicine. 2006;34:217-231.

Yang SW, Shen YC, Chen CH. Steroids and triterpenoids of Antrodia cinnamomea --a fungus parasitic on cinnamomum micranthum. Phytochemistry.1996;41:1389-1392.

Yao JS, Chen YM, Zhai WW,Young KW, Yang GY. Minocycline Exerts Multiple Inhibitory Effects on Vascular Endothelial Growth Factor-Induced Smooth Cell Migration. Circulation research. 2004;95:364-371.

Yokouchi K, Numaguchi Y, Kubota R, Ishii M, Imai H, Murakami R, Ogawa Y, Kondo T, Okumura K. l-Caldesmon Regulates Proliferation and Migration of Vascular Smooth Muscle Cells and Inhibits Neointimal Formation After Angioplasty. Arteriosclerosis, Thrombosis,and Vascular Biology. 2006;26:2231-2237.

Zempo N, Kenagy RD, Au YPT, et al. Matrix metalloproteinases of vascular wall cells are increased in balloon-injured rat carotid artery. Joural of Vascular Surgery. 1994;20: 209–217.
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