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研究生:陳建旭
研究生(外文):Chen,Chienhsu
論文名稱:天山雪蓮 Saussurea involucrate 萃取物及其純化物 Rutin 對抗老化作用之探討
論文名稱(外文):The investigation of the anti-aging mechanism of Saussureainvolucrate and Rutin
指導教授:陳怡伶陳怡伶引用關係
指導教授(外文):Chen,Yilin
口試委員:俞松良趙國評花國鋒李柏磊陳怡伶
口試委員(外文):Yu, SungliangChao, LouiskuopingHua, KuofengLee, Po-leiChen,Yilin
口試日期:2011-06-17
學位類別:碩士
校院名稱:國立宜蘭大學
系所名稱:生物技術研究所碩士班
學門:農業科學學門
學類:畜牧學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:中文
論文頁數:58
中文關鍵詞:黃酮類天山雪蓮芸香素第二型環氧酶抗老化
外文關鍵詞:FlavonoidSaussurea involucrateRutinCyclooxygenase-2Anti-agingpyknotic nuclei
相關次數:
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  • 下載下載:134
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在許多的神經退化性疾病都可以明顯的觀察到細胞中的抗氧化能力
失去平衡,並產生大量的自由基去氧化核酸、脂質及蛋白質,使得細胞
走向凋亡,若能夠透過保護細胞的抗氧化能力來降低細胞的凋亡,就能
夠避免神經退化性疾病的發生。
天山雪蓮 (Saussurea involucrate) 乙酸乙酯層萃取物在先前的研究
已發現具有對抗攝護腺癌細胞的生長,而在其萃取物中也發現主要成分
大多為黃酮類的物質,在許多的研究中都指出黃酮類 (flavonid) 具有良
好的抗氧化效果。另外,芸香素 (Rutin) 為天山雪蓮萃取物中的主要成
分之一,其也屬於黃酮類物質,並且能夠預防神經退化性疾病。
氧化壓力的提高與老化的過程有明顯關聯,因此本研究利用半乳糖
(D-galactose) 來模擬動物因氧化壓提高之老化體內模式,觀察天山雪蓮
萃取物及 Rutin 是否能夠保護神經細胞。由結果顯示,天山雪蓮萃取物
及 Rutin 可經由抑制脂質過氧化產物丙二醛 (MDA) 的表現,並活化穀
胱甘肽過氧化酶(glutathione peroxidase,GPx)及超氧化物歧化酶
(Superoxide Dismutase,SOD) 的活性,同時天山雪蓮乙酸乙酯層萃取
物及 Rutin 亦可透過抑制 NF-κB 的表現量來降低第二型環氧酶
(COX-2) 之蛋白質表現量,並且降低細胞凋亡蛋白 PARP 及 caspase-3
的表現,進而達到保護神經細胞之作用。
This study was carried out to investigate the protective effect of Saussurea
involucrate and Rutin against D-galactose (D-gal)-induced brain injury in mice. .
Protein expression was determined by Western blot. Apoptotic cells were determined
by H&E stain. D-galactose aging animal treatment was used as a model system to study
the anti-oxidant effects of Saussurea involucrate and Rutin in vivo.
Pretreatment with Saussurea involucrate and Rutin increased cell viability and
decreased membrane damage, generation of reactive oxygen species and degradation of
poly (ADP-ribose) polymerase in D-galactose-treated mice and also decreased the
expression of cyclooxygenase-2, via downregulation of NF-kappaB, resulting in a
decrease in lipid peroxidation. These results suggest that Saussurea involucrate and Rutin
are potential antioxidant agents. In another set of experiments, we evaluated the effect of
Saussurea involucrate and Rutin in a murine aging model, in which chronic systemic
exposure to D-galactose (D-gal) causes the acceleration of senescence. Administration of
Saussurea involucrate and Rutin (30 mg/kg/d and 30 mg/kg/d) for 6 weeks concomitant
with D-gal injection significantly increased superoxide dismutase and glutathione
peroxidase activities and decreased the MDA level in plasma and brain. Furthermore, we
used H&E staining to quantify cell death within hippocampus by counting the pyknotic
nuclei in H&E section. We found that percentage of pyknotic nuclei in the D-gal-treated
mice was much higher than in controls.These results suggest that Saussurea involucrate
and Rutin exert potent antiaging effects against D-gal in mice via antioxidative
mechanisms.
中文摘要……………………………………………………………………I
英文摘要……………………………………………………………………II
目錄………………………………………………………………………III
圖表目錄…………………………………………………………………V
縮寫表……………………………………………………………………VI
第一章 前言………………………………………………………………1
第一節 老化與氧化壓力…………………………………………………1
第二節 活性氧物質 (ROS) ……………………………………………4
第三節 抗氧化防禦系統…………………………………………………5
第四節 氧化壓力與腦部相關疾病………………………………………10
第五節 第二型環氧化酶與神經傷害……………………………………13
第六節 大腦海馬迴 (Hippocampus) ……………………………………16
第七節 半乳糖老化模式…………………………………………………18
第八節 天山雪蓮 Saussurea involucrate 與芸香素 Rutin……………19
第二章 實驗目的…………………………………………………………22
第三章 實驗材料…………………………………………………………23
第一節 實驗儀器…………………………………………………………23
第二節 藥品詴劑…………………………………………………………24
第四章 實驗方法…………………………………………………………27
第一節 實驗流程…………………………………………………………27
第二節 半乳糖老化模式…………………………………………………28
第三節 血漿樣本之處理…………………………………………………28
第四節 免疫組織染色……………………………………………………28
第五節 丙二醛含量之測量………………………………………………29
第六節 超氧化物岐化酵素活性測量……………………………………30
第七節 穀胱甘肽過氧化酶活性之測量…………………………………31
第八節 西方墨點法………………………………………………………33
第五章 實驗結果…………………………………………………………34
第一節 小鼠體重增加情形………………………………………………34
第二節 半乳糖老化小鼠血漿中老化生理指標分析……………………34
第三節大腦 PARP 及 Caspase-3 表現情形……………………………34
第四節 環氧酶及其上游轉錄因子之表現量………………………………35
第五節 大腦海馬迴組織 cleaved caspase-3 表現情形…………………35
第六章 討論………………………………………………………………37
第七章 結論與未來展望…………………………………………………41
第八章 結果圖表…………………………………………………………42
參考文獻……………………………………………………………………51
1. Barnes, C.A. (1979). Memory deficits associated with senescence: a
neurophysiological and behavioral study in the rat. J. Comp. Physiol.
Psychol. 93, 74–104.
2. Basselin, M., Chang, L., Chen, M., Bell, J.M., and Rapoport, S.I. (2008).
Chronic administration of valproic acid reduces brain NMDA signaling
via arachidonic acid in unanesthetized rats. Neurochem Res 33,
2229-2240.
3. Baynes, W.J. (2001). The role of AGEs in aging: causation or correlation.
Expt. Geront. 36, 1527–1537.
4. Burton, G.W., and Ingold, K.U. (1984). beta-Carotene: an unusual type
of lipid antioxidant. Science 224, 569-573.
5. Butterfield, D.A., and Lauderback, C.M. (2002). Lipid peroxidation and
protein oxidation in Alzheimers disease brain: potential causes and
consequences involving amyloid beta-peptide-associated free radical
oxidative stress. Free Radic Biol Med 32, 1050-1060.
6. Calleja, M., Pena, P., Ugalde, C., Ferreiro, C., Marco, R., and Garesse, R.
(1993). Mitochondrial DNA remains intact during Drosophila aging, but
the levels of mitochondrial transcripts are significantly reduced. J Biol
Chem 268, 18891-18897.
7. Cameron, E., and Pauling, L. (1976). Supplemental ascorbate in the
supportive treatment of cancer: Prolongation of survival times in
terminal human cancer. Proc Natl Acad Sci U S A 73, 3685-3689.
8. Carr, A., and Frei, B. (1999). Does vitamin C act as a pro-oxidant under
physiological conditions? FASEB J 13, 1007-1024.
9. Cecchi, C., Latorraca, S., Sorbi, S., Iantomasi, T., Favilli, F., Vincenzini,
M.T., and Liguri, G. (1999). Gluthatione level is altered in lymphoblasts
from patients with familial Alzheimers disease. Neurosci Lett 275,
152-154.
10. Choi, S.H., Aid, S., and Bosetti, F. (2009). The distinct roles of
cyclooxygenase-1 and -2 in neuroinflammation: implications for
translational research. Trends Pharmacol Sci 30, 174-181.
11. Cui, X., Zuo, P., Zhang, Q., Li, X., Hu, Y., Long, J., Packer, L., and Liu,
J. (2006). Chronic systemic D-galactose exposure induces memory loss,
neurodegeneration, and oxidative damage in mice: protective effects of
R-alpha-lipoic acid. J Neurosci Res 84, 647-654.
12. Culmsee, C., and Krieglstein, J. (2007). Ischaemic brain damage after
stroke: new insights into efficient therapeutic strategies. International
Symposium on Neurodegeneration and Neuroprotection. EMBO Rep 8,
129-133.
13. Cruz, T., Galvez, J., Ocete, M.A., Crespo, M.E., Sanchez de Medina
LHF., and Zarzuelo, A. (1998). Oral administration of rutoside can
ameliorate inflammatory bowel disease in rats. Life Sci 62, 687–695.
14. Del Rio, P., Montiel, T., Chagoya, V., and Massieu, L. (2007).
Exacerbation of excitotoxic neuronal death induced during
mitochondrial inhibition in vivo: relation to energy imbalance or ATP
depletion? Neuroscience 146, 1561-1570.
15. Emerit, J., Edeas, M., and Bricaire, F. (2004). Neurodegenerative
diseases and oxidative stress. Biomed Pharmacother 58, 39-46.
16. Feng, L., Xia, Y., Garcia, G.E., Hwang, D., and Wilson, C.B. (1995).
Involvement of reactive oxygen intermediates in cyclooxygenase-2
expression induced by interleukin-1, tumor necrosis factor-alpha, and
lipopolysaccharide. J Clin Invest 95, 1669-1675.
17. Finkel, T., and Holbrook, N.J. (2000). Oxidants, oxidative stress and the
biology of ageing. Nature 408, 239-247.
18. Flanary, B.E., and Streit, W.J. (2006). Alpha-tocopherol (vitamin E)
induces rapid, nonsustained proliferation in cultured rat microglia. Glia
53, 669-674.
19. Floyd, R., and Hensley, K., (2002). Oxidative stress in brain aging
implications for therapeutics of neurodegenerative diseases. Neurobiol.
Aging 23, 795–807.
20. Guardia, T., Rotelli, A.E., Juarez, A.O., and Pelzer, L.E. (2001).
Anti-inflammatory properties of plant flavonoids. Effects of rutin,
quercetin and hesperidin on adjuvant arthritis in rat. Farmaco
56,683-687.
21. Gupta, R., Singh, M., and Sharma, A. (2003). Neuroprotective effect of
antioxidants on ischaemia and reperfusion-induced cerebral injury.
Pharmacol Res 48,209-215.
22. Gustafsson, A.B., Sayen, M.R., Williams, S.D., Crow, M.T., and
Gottlieb, R.A. (2002). TAT protein transduction into isolated perfused
hearts: TAT-apoptosis repressor with caspase recruitment domain is
cardioprotective. Circulation 106, 735-739.
23. Harman, D. (1956). Aging: a theory based on free radical and radiation
chemistry. J Gerontol 11, 298-300.
24. Hensley, K., Carney, J.M., Mattson, M.P., Aksenova, M., Harris, M.,
Wu, J.F., Floyd, R.A., and Butterfield, D.A. (1994). A model for
beta-amyloid aggregation and neurotoxicity based on free radical
generation by the peptide: relevance to Alzheimer disease. Proc Natl
Acad Sci U S A 91, 3270-3274.
25. Hewett, S.J., Uliasz, T.F., Vidwans, A.S., and Hewett, J.A. (2000).
Cyclooxygenase-2 contributes to N-methyl-D-aspartate-mediated
neuronal cell death in primary cortical cell culture. J Pharmacol Exp
Ther 293, 417-425.
26. Hsieh, H., Wua, W., and Hu, M. (2009). Soy isoflavones attenuate
oxidative stress and improve parameters related to aging and
Alzheimer’s disease in C57BL/6J mice treated with D-galactose. Food
Chem. Toxicol. 47, 625–632.
27. Hunsaker, M.R., Lee, B., and Kesner, R.P. (2008). Evaluating the
temporal context of episodic memory: the role of CA3 and CA1. Behav
Brain Res 188:310–315.
28. Im, J.Y., Kim, D., Paik, S.G., and Han, P.L. (2006).
Cyclooxygenase-2-dependent neuronal death proceeds via superoxide
anion generation. Free Radic Biol Med 41, 960-972.
29. Ikawa, H., Kameda, H., Kamitani, H., Baek, S.J., Nixon, J.B., Hsi, L.C.,
and Eling, T.E. (2001). Effect of PPAR activators on
cytokine-stimulated cyclooxygenase-2 expression in human colorectal
carcinoma cells. Exp Cell Res 267, 73-80.
30. Jang, Y.C., and Remmen, H.V. (2009). The mitochondrial theory of
aging: insight from transgenic and knockout mouse models. Exp.
Gerontol. 44, 256–260.
31. Jones, D.P., Carlson, J.L., Mody, V.C., Cai, J., Lynn, M.J., and
Sternberg, P. (2000). Redox state of glutathione in human plasma. Free
Radic Biol Med 28, 625-635.
32. Kaplan, L.A., and Pesce, A.J. (1996). Clinical Chemistry, third ed.
Mosby, St. Louis. p. 954.
33. Kim, H., Kong, H., Choi, B., Yang, Y., Kim, Y., Lim, M.J., Neckers, L.,
and Jung, Y. (2005). Metabolic and pharmacological properties of rutin,
a dietary quercetin glycoside, for treatment of inflammatory bowel
disease. Pharm Res 22:1499–1509.
34. Kukreja, R.C., Kontos, H.A., Hess, M.L., and Ellis, E.F. (1986). PGH
synthase and lipoxygenase generate superoxide in the presence of
NADH or NADPH. Circ Res 59, 612-619.
35. Lu, J., Xing, J., and Li, J. (2007). Prostaglandin E2 (PGE2) inhibits
glutamatergic synaptic transmission in dorsolateral periaqueductal gray
(dl-PAG). Brain Res 1162, 38-47.
36. Masella, R., Di Benedetto, R., Vari, R., Filesi, C., and Giovannini, C.
(2005). Novel mechanisms of natural antioxidant compounds in
biological systems: involvement of glutathione and glutathione-related
enzymes. J Nutr Biochem 16, 577-586.
37. Morris, R.G., Garrud, P., Rawlins, J.N., and O’Keefe, J. (1982). Place
navigation impaired in rats with hippocampal lesions. Nature
297:681–683.
38. McCarty, M.F. (2006). Toward prevention of Alzheimers
disease--potential nutraceutical strategies for suppressing the production
of amyloid beta peptides. Med Hypotheses 67, 682-697.
39. McCord, J.M., and Fridovich, I. (1969). Superoxide dismutase. An
enzymic function for erythrocuprein (hemocuprein). J Biol Chem 244,
6049-6055.
40. Miettinen, S., Fusco, F.R., Yrjanheikki, J., Keinanen, R., Hirvonen, T.,
Miller, D.M., Buettner, G.R., and Aust, S.D. (1990). Transition metals as
catalysts of "autoxidation" reactions. Free Radic Biol Med 8, 95-108.
41. Miquel, J., Economos, A.C., Fleming, J., and Johnson, Jr., J.E. (1980).
Mitochondrial role in cell aging. Exp. Gerontol. 15, 575-591.
42. Nakamura, T., and Sakamoto, K. (2001). Reactive oxygen species
up-regulates cyclooxygenase-2, p53, and Bax mRNA expression in
bovine luteal cells. Biochem Biophys Res Commun 284, 203-210.
43. Nogueira, C.W., Zeni, G., and Rocha, J.B. (2004). Organoselenium and
organotellurium compounds: toxicology and pharmacology. Chem Rev
104, 6255-6285.
44. Richetti, S.K., Blank, M., Capiotti, K.M., Piato A.L., Bogo, M.R.,
Vianna, M.R., and Bonan, C.D. (2011). Quercetin and rutin prevent
scopolamine-induced memory impairment in zebrafish. Behav Brain Res
217, 10-5.
45. Rockwell, P., Martinez, J., Papa, L., and Gomes, E. (2004). Redox
regulates COX-2 upregulation and cell death in the neuronal response to
cadmium. Cell Signal 16, 343-353.
46. Rosenzweig, E.S., and Barnes, C.A. (2003). Impact of aging on
hippocampal function: plasticity, network dynamics, and cognition. Prog.
Neurobiol. 69, 143-179.
47. Ryter, S.W., and Tyrrell, R.M. (1998). Singlet molecular oxygen ((1)O2):
a possible effector of eukaryotic gene expression. Free Radic Biol Med
24, 1520-1534.
48. Sanzgiri, R.P., Araque, A., and Haydon, P.G. (1999). Prostaglandin E(2)
stimulates glutamate receptor-dependent astrocyte neuromodulation in
cultured hippocampal cells. J Neurobiol 41, 221-229.
49. Shen, D., Dalton, T.P., Nebert, D.W., and Shertzer, H.G. (2005).
Glutathione redox state regulates mitochondrial reactive oxygen
production. J Biol Chem 280, 25305-25312.
50. Shim, M., and Eling, T.E. (2008). Vitamin E succinate induces NAG-1
expression in a p38 kinase-dependent mechanism. Mol Cancer Ther 7,
961-971.
51. Song, X., Bao, M., Li, D., and Li, Y. (1999). Advanced glycation in
D-galactose induced mouse aging model. Mech. Ageing Dev. 108,
239–251.
52. Stella, N., Tence, M., Glowinski, J., and Premont, J. (1994).
Glutamate-evoked release of arachidonic acid from mouse brain
astrocytes. J Neurosci 14, 568-575.
53. Suh, J.H., Moreau, R., Heath, S.H., and Hagen, T.M. (2005). Dietary
supplementation with (R)-alpha-lipoic acid reverses the age-related
accumulation of iron and depletion of antioxidants in the rat cerebral
cortex. Redox Rep 10, 52-60.
54. Taylor, A.L., and Hewett, S.J. (2002). Potassium-evoked glutamate
release liberates arachidonic acid from cortical neurons. J Biol Chem
277, 43881-43887.
55. Tomoko, K., Yoshiki, K., and Hideki, I. (2008). Protective effect of rutin
against spatial memory impairment induced by trimethyltin in rats.
Nutrition Research 28,629-634.
56. Tzeng, S.F., Hsiao, H.Y., and Mak, O.T. (2005). Prostaglandins and
cyclooxygenases in glial cells during brain inflammation. Curr Drug
Targets Inflamm Allergy 4, 335-340.
57. Valko, M., Leibfritz, D., Moncol, J., Cronin, M.T., Mazur, M., and
Telser, J. (2007). Free radicals and antioxidants in normal physiological
functions and human disease. Int J Biochem Cell Biol 39, 44-84.
58. Valko, M., Morris, H., and Cronin, M.T. (2005). Metals, toxicity and
oxidative stress. Curr Med Chem 12, 1161-1208.
59. Valko, M., Rhodes, C.J., Moncol, J., Izakovic, M., and Mazur, M.
(2006). Free radicals, metals and antioxidants in oxidative stress-induced
cancer. Chem Biol Interact 160, 1-40.
60. Wang, Q., Tang, X.N., and Yenari, M.A. (2007). The inflammatory
response in stroke. J Neuroimmunol 184, 53-68.
61. Way, T.D., Lee, J.C., Kuo, D.H., Fan, L.L., Huang, C.H., Lin, H.Y.,
Shieh, P.C., Kuo, P.T., Liao, C.F., Liu, H., and Kao, J.Y. (2010)
Inhibition of epidermal growth factor receptor signaling by Saussurea
involucrata, a rare traditional Chinese medicinal herb, in human
hormone-resistant prostate cancer PC-3 cells. J Agric Food Chem 58,
3356–3365.
62. Yang, H., and Chen, C. (2008). Cyclooxygenase-2 in synaptic signaling.
Curr Pharm Des 14, 1443-1451.
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