跳到主要內容

臺灣博碩士論文加值系統

(44.200.171.74) 您好!臺灣時間:2022/08/12 07:07
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

: 
twitterline
研究生:呂政澤
研究生(外文):Chen-Che Lu
論文名稱:牛樟芝純化成分抑制幽門螺旋桿菌誘導人類胃上皮細胞的發炎作用
論文名稱(外文):Antrodia camphorata constituents inhibit Helicobacter pylori- induced inflammation in human gastric epithelial cells
指導教授:賴志河
指導教授(外文):Chih-Ho Lai
學位類別:碩士
校院名稱:中國醫藥大學
系所名稱:基礎醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:英文
論文頁數:45
中文關鍵詞:幽門螺旋菌牛樟芝發炎作用
外文關鍵詞:H. pyloriAntrodia camphoratainflammationNF-κBIL-8
相關次數:
  • 被引用被引用:4
  • 點閱點閱:2083
  • 評分評分:
  • 下載下載:1
  • 收藏至我的研究室書目清單書目收藏:1
幽門螺旋菌 (H. pylori) 是一種微好氧螺旋形狀的革蘭氏陰性菌,會存在於人類的胃以及十二指腸的表皮黏膜上。幽門螺旋菌會引起胃襯的慢性發炎並且與十二指腸、胃潰瘍和以及胃癌的發展有密切關係。幽門螺旋菌對於胃腸道潰瘍的發生有密切相關,一般認為滅除幽門螺旋菌可以使得胃發炎復原,亦可治療潰瘍性疾病。現有多種治療幽門螺旋菌感染的療法,最為普遍使用的是三合一療法,包含了兩種抗生素與一種胃酸分泌抑制劑。然而,在治療幽門螺旋菌感染上,抗生素抗藥性的增加與抗微生物劑的使用有關。因此,根治幽門螺旋菌過程中需要有輔助治療的方法。樟薄孔菌(Antrodia camphorata)是在台灣的一種稀有的藥材,其子果實的名稱是眾所周知的牛樟芝或樟芝。在台灣,牛樟芝對人體的神奇功效,被許多機構廣泛的調查與研究。在本篇論文中,從牛樟芝上分離出13種萃取物。在13種萃取物中,不論是在胃上皮細胞與小鼠巨噬細胞上,AC-3 (15α-Acetyldehydrosulphurenic acid)與AC-10 (Antcin C)對於幽門螺旋菌擁有較高的抑制效果。此外,對於NF-κB以及IL-8的抑制,也顯示出AC萃取物的抗發炎功效。期望此實驗結果可以為治療幽門螺旋菌感染帶來新的方法。
Helicobacter pylori is a Gram-negative, micro-aerophilic bacterium that inhabits various areas of the stomach and duodenum. It causes a chronic inflammation of the stomach lining and strongly linked to the development of duodenal and gastric ulcers and stomach cancer. It has been well confirmed that infection with H. pylori is an etiological agent of peptic ulcer, and eradication of this organism can not only make the associated gastritis regression but cure these diseases. There are several treatments of H. pylori infection, most common treatment is triple therapy, including two antibiotics and a proton pump inhibitor. However, the increasing of antibiotic resistant rate was associated with the usage of antimicrobial agents in the treatment of H. pylori infection. Thus, the complementary therapeutic methods in eradication of H. pylori were needed. Antrodia camphorata (A. camphorata) is a rare treasure in Taiwan, the fruiting body of A. camphorata is well known by named niu-chang-chih or jang-jy. The miraculous efficacy in human body had been investigated and researched by many organizations in Taiwan. In this study, thirteen compounds were isolated from A. camphorata. Among them, compound 15α-Acetyldehydrosulphurenic acid (number 3) and Antcin C (number 10) had higher inhibition effects on H. pylori-induced inflammatory responses in gastric epithelial cells and mouse macrophage RAW 264.7 cells than other compounds. In addition, these constituents also inhibited the IL-8 protein expression and NF-κB signal activation in H. pylori-infected gastric epithelial cells and it might contribute to the potential role of A. camphorata as an anti-inflammatory action. Results from our study might bring a new method for treating of H. pylori infection.
Content
Abstract III
中文摘要 (Abstract in Chinese) V
Introduction 1
Helicobacter pylori infection and inflammation responses 1
Treatments of H. pylori infection 2
Antrodia camphorata 5
Materials and methods 8
Bacterial strain and cell culture 8
Determination of anti-bacterial activity 8
Cell survival assay (MTT method) 9
Inhibition of H. pylori adherence to and invasion into epithelial cells 9
Transient transfection of NF-κB and IL-8 reporter genes 10
Measurement of interleukin-8 expression 11
Inhibition of macrophage activation 11
Western blot for assay of iNOS expression 12
Cell survival assay (MTT method) in macrophages 12
Statistical evaluation 13
Results 14
Inhibition of H. pylori growth 14
Treatment of A. camphorata constituents AC-3, AC-10, and AC-12 does influence epithelial cell viability 14
Inhibition of H. pylori adherence and invasion of gastric epithelial cells 15
AC-3 inhibit H. pylori-induced inflammation of gastric epithelial cells 15
Treatment of A. camphorata constituents AC-3, AC-10, and AC-12 does influence marcophages viability 17
A. camphorata constituents AC-3 and AC-10 inhibit LPS-mediated nitric oxide production in macrophages 17
Western blot of inhibition of iNOS production 18
Discussion and conclusion 19
References 22
Table 1 Molecular weight of A. camphorata constituents 30
Table 2 Inhibitory effects of A. camphorata constituents 31
Fig. 1. Chemiacl structures of A. camphorata constituents 1-11 32
Fig. 2. Effect of A. camphorata constituents (AC-3, AC-10, and AC-12) and DMSO control on gastric epithelial cell viability 35
Fig. 3. Effect of A. camphorata constituents (AC-3, AC-10, and AC-12) on H. pylori adhesion of gastric epithelial cells 36
Fig. 4. Effect of A. camphorata constituents (AC-3, AC-10, and AC-12) on H. pylori invasion of gastric epithelial cells 37
Fig. 5. Effect of A. camphorata constituents (AC-3, AC-10, and AC-12) on H. pylori-induced NF-κB activation in gastric epithelial cells 38
Fig. 6. Effect of A. camphorata constituents (AC-3, AC-10, and AC-12) on H. pylori-induced IL-8–luciferase activity in gastric epithelial cells. 39
Fig. 7. Effect of A. camphorata constituents (AC-3, AC-10, and AC-12) and DMSO control in RAW 264.7 cells 40
Fig. 8. Inhibition by A. camphorata constituents (AC-3, AC-10, and AC-12) and DMSO control of LPS-induced NO production in RAW 264.7 cells 41
Fig. 9. Effects of A. camphorata constituents (AC-3 and AC-10) on the iNOS protein expression in RAW 264.7 cells 42
Appendix 1 Therapy of H. pylori infection 43
Appendix 2 Drugs used in H. pylori therapy 44
Ao, Z.H., Xu, Z.H., Lu, Z.M., Xu, H.Y., Zhang, X.M. and Dou, W.F., 2009. Niuchangchih (Antrodia camphorata) and its potential in treating liver diseases. J Ethnopharmacol 121, 194-212.
Bochenek, W.J., Peters, S., Fraga, P.D., Wang, W., Mack, M.E., Osato, M.S., El-Zimaity, H.M., Davis, K.D. and Graham, D.Y., 2003. Eradication of Helicobacter pylori by 7-day triple-therapy regimens combining pantoprazole with clarithromycin, metronidazole, or amoxicillin in patients with peptic ulcer disease: results of two double-blind, randomized studies. Helicobacter 8, 626-642.
Brandt, S., Kwok, T., Hartig, R., Konig, W. and Backert, S., 2005. NF-kappaB activation and potentiation of proinflammatory responses by the Helicobacter pylori CagA protein. Proc Natl Acad Sci U S A 102, 9300-9305.
Chan, Y. and Chan, C.H., 2003. Antibiotic resistance of pathogenic bacteria from odontogenic infections in Taiwan. J Microbiol Immunol Infect 36, 105-110.
Chen, K.C., Peng, C.C., Peng, R.Y., Su, C.H., Chiang, H.S., Yan, J.H. and Hsieh-Li, H.M., 2007. Unique formosan mushroom Antrodia camphorata differentially inhibits androgen-responsive LNCaP and -independent PC-3 prostate cancer cells. Nutr Cancer 57, 111-121.
Chien, S.C., Chen, M.L., Kuo, H.T., Tsai, Y.C., Lin, B.F. and Kuo, Y.H., 2008. Anti-inflammatory activities of new succinic and maleic derivatives from the fruiting body of Antrodia camphorata. J Agric Food Chem 56, 7017-7022.
Chourasia, D., Misra, A., Pandey, R. and Ghoshal, U.C., 2008. Gastric atrophy and intestinal metaplasia in a patient on long-term proton pump inhibitor therapy. Trop Gastroenterol 29, 172-174.
de Boer, W.A., Driessen, W.M., Jansz, A.R. and Tytgat, G.N., 1995. Quadruple therapy compared with dual therapy for eradication of Helicobacter pylori in ulcer patients: results of a randomized prospective single-centre study. Eur J Gastroenterol Hepatol 7, 1189-1194.
de Boer, W.A., Driessen, W.M., Potters, V.P. and Tytgat, G.N., 1994. Randomized study comparing 1 with 2 weeks of quadruple therapy for eradicating Helicobacter pylori. Am J Gastroenterol 89, 1993-1997.
Filipec Kanizaj, T., Katicic, M., Skurla, B., Ticak, M., Plecko, V. and Kalenic, S., 2009. Helicobacter pylori eradication therapy success regarding different treatment period based on clarithromycin or metronidazole triple-therapy regimens. Helicobacter 14, 29-35.
Fong, Y.C., Maa, M.C., Tsai, F.J., Chen, W.C., Lin, J.G., Jeng, L.B., Yang, R.S., Fu, W.M. and Tang, C.H., 2008. Osteoblast-derived TGF-beta1 stimulates IL-8 release through AP-1 and NF-kappaB in human cancer cells. J Bone Miner Res 23, 961-970.
Gobert, A.P., McGee, D.J., Akhtar, M., Mendz, G.L., Newton, J.C., Cheng, Y., Mobley, H.L. and Wilson, K.T., 2001. Helicobacter pylori arginase inhibits nitric oxide production by eukaryotic cells: a strategy for bacterial survival. Proc Natl Acad Sci U S A 98, 13844-13849.
Graham, D.Y., Lu, H. and Yamaoka, Y., 2009. African, Asian or Indian enigma, the East Asian Helicobacter pylori: facts or medical myths. J Dig Dis 10, 77-84.
Gu, Q., Xia, H.H., Wang, J.D., Wong, W.M., Chan, A.O., Lai, K.C., Chan, C.K., Yuen, M.F., Fung, F.M., Wong, K.W., Lam, S.K. and Wong, B.C., 2006. Update on clarithromycin resistance in Helicobacter pylori in Hong Kong and its effect on clarithromycin-based triple therapy. Digestion 73, 101-106.
Hacin, B., Rogelj, I. and Matijasic, B.B., 2008. Lactobacillus isolates from weaned piglets'' mucosa with inhibitory activity against common porcine pathogens. Folia Microbiol (Praha) 53, 569-576.
Harris, P.R., Smythies, L.E., Smith, P.D. and Dubois, A., 2000. Inflammatory cytokine mRNA expression during early and persistent Helicobacter pylori infection in nonhuman primates. J Infect Dis 181, 783-786.
Hentschel, E., Brandstatter, G., Dragosics, B., Hirschl, A.M., Nemec, H., Schutze, K., Taufer, M. and Wurzer, H., 1993. Effect of ranitidine and amoxicillin plus metronidazole on the eradication of Helicobacter pylori and the recurrence of duodenal ulcer. N Engl J Med 328, 308-312.
Hseu, Y.C., Chen, S.C., Yech, Y.J., Wang, L. and Yang, H.L., 2008. Antioxidant activity of Antrodia camphorata on free radical-induced endothelial cell damage. J Ethnopharmacol 118, 237-245.
Hsiao, G., Shen, M.Y., Lin, K.H., Lan, M.H., Wu, L.Y., Chou, D.S., Lin, C.H., Su, C.H. and Sheu, J.R., 2003. Antioxidative and hepatoprotective effects of Antrodia camphorata extract. J Agric Food Chem 51, 3302-3308.
Hsu, P.I., Lai, K.H., Lin, C.K., Chen, W.C., Yu, H.C., Cheng, J.S., Tsay, F.W., Wu, C.J., Lo, C.C., Tseng, H.H., Yamaoka, Y., Chen, J.L. and Lo, G.H., 2005. A prospective randomized trial of esomeprazole- versus pantoprazole-based triple therapy for Helicobacter pylori eradication. Am J Gastroenterol 100, 2387-2392.
Jodlowski, T.Z., Lam, S. and Ashby, C.R., Jr., 2008. Emerging therapies for the treatment of Helicobacter pylori infections. Ann Pharmacother 42, 1621-1639.
Kim, S.Y., Lee, Y.C., Kim, H.K. and Blaser, M.J., 2006. Helicobacter pylori CagA transfection of gastric epithelial cells induces interleukin-8. Cell Microbiol 8, 97-106.
Lai, C.H., Chang, Y.C., Du, S.Y., Wang, H.J., Kuo, C.H., Fang, S.H., Fu, H.W., Lin, H.H., Chiang, A.S. and Wang, W.C., 2008a. Cholesterol depletion reduces Helicobacter pylori CagA translocation and CagA-induced responses in AGS cells. Infect Immun 76, 3293-3303.
Lai, C.H., Fang, S.H., Rao, Y.K., Geethangili, M., Tang, C.H., Lin, Y.J., Hung, C.H., Wang, W.C. and Tzeng, Y.M., 2008b. Inhibition of Helicobacter pylori-induced inflammation in human gastric epithelial AGS cells by Phyllanthus urinaria extracts. J Ethnopharmacol 118, 522-526.
Lai, C.H., Kuo, C.H., Chen, P.Y., Poon, S.K., Chang, C.S. and Wang, W.C., 2006. Association of antibiotic resistance and higher internalization activity in resistant Helicobacter pylori isolates. J Antimicrob Chemother 57, 466-471.
Lam, S.K. and Talley, N.J., 1998. Report of the 1997 Asia Pacific Consensus Conference on the management of Helicobacter pylori infection. J Gastroenterol Hepatol 13, 1-12.
Lee, I.H., Huang, R.L., Chen, C.T., Chen, H.C., Hsu, W.C. and Lu, M.K., 2002. Antrodia camphorata polysaccharides exhibit anti-hepatitis B virus effects. FEMS Microbiol Lett 209, 63-67.
Logan, R.P., Gummett, P.A., Misiewicz, J.J., Karim, Q.N., Walker, M.M. and Baron, J.H., 1991. One week eradication regimen for Helicobacter pylori. Lancet 338, 1249-1252.
Lu, Z.M., Tao, W.Y., Zou, X.L., Fu, H.Z. and Ao, Z.H., 2007. Protective effects of mycelia of Antrodia camphorata and Armillariella tabescens in submerged culture against ethanol-induced hepatic toxicity in rats. J Ethnopharmacol 110, 160-164.
Marshall, B., 2002. Helicobacter pylori: 20 years on. Clin Med 2, 147-152.
McMahon, B.J., Hennessy, T.W., Bensler, J.M., Bruden, D.L., Parkinson, A.J., Morris, J.M., Reasonover, A.L., Hurlburt, D.A., Bruce, M.G., Sacco, F. and Butler, J.C., 2003. The relationship among previous antimicrobial use, antimicrobial resistance, and treatment outcomes for Helicobacter pylori infections. Ann Intern Med 139, 463-469.
Megraud, F. and Lamouliatte, H., 2003. Review article: the treatment of refractory Helicobacter pylori infection. Aliment Pharmacol Ther 17, 1333-1343.
Mirbagheri, S.A., Hasibi, M., Abouzari, M. and Rashidi, A., 2006. Triple, standard quadruple and ampicillin-sulbactam-based quadruple therapies for H. pylori eradication: a comparative three-armed randomized clinical trial. World J Gastroenterol 12, 4888-4891.
Nozawa, Y., Nishihara, K., Peek, R.M., Nakano, M., Uji, T., Ajioka, H., Matsuura, N. and Miyake, H., 2002. Identification of a signaling cascade for interleukin-8 production by Helicobacter pylori in human gastric epithelial cells. Biochem Pharmacol 64, 21-30.
Parsonnet, J., 1998. Helicobacter pylori. Infect Dis Clin North Am 12, 185-197.
Peek, R.M., 2004. Helicobacter pylori and Gastroesophageal Reflux Disease. Curr Treat Options Gastroenterol 7, 59-70.
Poon, S.K., Chang, C.S., Su, J., Lai, C.H., Yang, C.C., Chen, G.H. and Wang, W.C., 2002. Primary resistance to antibiotics and its clinical impact on the efficacy of Helicobacter pylori lansoprazole-based triple therapies. Aliment Pharmacol Ther 16, 291-296.
Rao, Y.K., Fang, S.H. and Tzeng, Y.M., 2007. Evaluation of the anti-inflammatory and anti-proliferation tumoral cells activities of Antrodia camphorata, Cordyceps sinensis, and Cinnamomum osmophloeum bark extracts. J Ethnopharmacol 114, 78-85.
Salcedo, J.A. and Al-Kawas, F., 1998. Treatment of Helicobacter pylori infection. Arch Intern Med 158, 842-851.
Sartor, R.B., 1994. Cytokines in intestinal inflammation: pathophysiological and clinical considerations. Gastroenterology 106, 533-539.
Shen, C.C., Kuo, Y.C. and Huang, R.L., 2003. New ergostane and lanostane from Antrodia camphorata. J Chin Med 14, 247-258.
Suerbaum, S. and Michetti, P., 2002. Helicobacter pylori infection. N Engl J Med 347, 1175-1186.
Tang, C.H., Yang, R.S., Chen, Y.F. and Fu, W.M., 2007. Basic fibroblast growth factor stimulates fibronectin expression through phospholipase C gamma, protein kinase C alpha, c-Src, NF-kappaB, and p300 pathway in osteoblasts. J Cell Physiol 211, 45-55.
Tankovic, J., Lamarque, D., Lascols, C., Soussy, C.J. and Delchier, J.C., 2001a. Clarithromycin resistance of Helicobacter pylori has a major impact on the efficacy of the omeprazole-amoxicillin-clarithromycin therapy. Pathol Biol (Paris) 49, 528-533.
Tankovic, J., Lamarque, D., Lascols, C., Soussy, C.J. and Delchier, J.C., 2001b. Impact of Helicobacter pylori resistance to clarithromycin on the efficacy of the omeprazole-amoxicillin-clarithromycin therapy. Aliment Pharmacol Ther 15, 707-713.
Walsh, C., 2000. Molecular mechanisms that confer antibacterial drug resistance. Nature 406, 775-781.
Walsh, J.H. and Peterson, W.L., 1995. The treatment of Helicobacter pylori infection in the management of peptic ulcer disease. N Engl J Med 333, 984-991.
Warren, J.R., 1983. Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet i, 1273.
Wong, W.M., Wong, B.C., Lu, H., Gu, Q., Yin, Y., Wang, W.H., Fung, F.M., Lai, K.C., Xia, H.H., Xiao, S.D. and Lam, S.K., 2002. One-week omeprazole, furazolidone and amoxicillin rescue therapy after failure of Helicobacter pylori eradication with standard triple therapies. Aliment Pharmacol Ther 16, 793-798.
Yamaoka, Y., Kita, M., Kodama, T., Sawai, N. and Imanishi, J., 1996. Helicobacter pylori cagA gene and expression of cytokine messenger RNA in gastric mucosa. Gastroenterology 110, 1744-1752.
Yang, H.L., Chen, C.S., Chang, W.H., Lu, F.J., Lai, Y.C., Chen, C.C., Hseu, T.H., Kuo, C.T. and Hseu, Y.C., 2006. Growth inhibition and induction of apoptosis in MCF-7 breast cancer cells by Antrodia camphorata. Cancer Lett 231, 215-227.
Zullo, A., Gatta, L., De Francesco, V., Hassan, C., Ricci, C., Bernabucci, V., Cavina, M., Ierardi, E., Morini, S. and Vaira, D., 2005. High rate of Helicobacter pylori eradication with sequential therapy in elderly patients with peptic ulcer: a prospective controlled study. Aliment Pharmacol Ther 21, 1419-1424.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關期刊