跳到主要內容

臺灣博碩士論文加值系統

(44.192.38.49) 您好!臺灣時間:2023/02/09 14:35
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:林佩璇
研究生(外文):Pei-Hsuan Lin
論文名稱:Hydroxyzine對P-glycoprotein作用之研究
指導教授:蘇聖芳蘇聖芳引用關係
指導教授(外文):Sheng-Fang Su
學位類別:碩士
校院名稱:國立成功大學
系所名稱:臨床藥學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:84
外文關鍵詞:hydroxyzineP-glycoprotein
相關次數:
  • 被引用被引用:5
  • 點閱點閱:426
  • 評分評分:
  • 下載下載:27
  • 收藏至我的研究室書目清單書目收藏:0
中文摘要
P-glycoprotein是多重抗藥性(MDR)基因的protein產物,且為一
efflux transporter,會降低藥物在細胞內的蓄積,因此被視為化學治療上的障礙。之前實驗已證實一結構與已知為Pgp抑制劑terfenadine相似的藥物hydroxyzine可增加etoposide在老鼠小腸中的吸收,但由於老鼠小腸中尚有其他transporters的存在,所以本實驗想更進一步探討hydroxyzine在Pgp上所扮演的角色。
本研究首先是建立一具有Pgp表現的HEK293(human embryonic kidney cell line)細胞株,再以不同的Pgp受質(vincristine, epirubicin, etoposide)進行細胞毒性試驗,得知在有Pgp表現的HEK293細胞株中EC50會明顯的增加。同時,藉由已被證實為Pgp受質的rhodamine123這個螢光性物質,來確認Pgp在HEK293細胞株的功能及表現,結果在有Pgp表現的HEK293細胞株中,rhodamine123的蓄積會降低;接著,再以rhodamine123當受質,以不同濃度的hydroxyzine當抑制劑,結果卻顯示在有MDR-transfected的HEK293 cells中 hydroxyzine並不會增加rhodamine123的蓄積。但由於不同的受質作用於Pgp上的位置並不完全相似,所以我們進一步再利用作用於另一個結合位置(H site)的受質colchicine來探討其與hydroxyzine之間的交互作用,結果顯示,hydroxyzine同樣也不會增加colchicine在MDR-transfected HEK293細胞中的蓄積,由以上結果可證實hydroxyzine並不會作用於Pgp來增加藥物在細胞中的蓄積。
Abstract
P-glycoprotein functions as an ATP-dependent conjugate efflux pump to decrease drug accumulation in a variety of systems. Therefore, it was regarded as a major obstacle in chemotherapy. Previous study has demonstrated that hydroxyzine could increase the uptake of etoposide in the rat intestine. Since there are other transporters existing in the rat intestine we further want to investigate the interaction between hydroxyzine and Pgp.
We have established the MDR-transfected HEK293 cell. The permanent expression of human MDR in stable transfected cell lines enabled us to study on cytotoxicity using Pgp substrates (vincristine, epirubicin, etoposide). Increased EC50 was observed in MDR-transfected HEK293 cells. Moreover, the function and expression of Pgp in HEK293 cells was confirmed using Pgp substrate R123. The accumulation of R123 was significantly decreased in MDR-transfected HEK293 cells. Furthermore, R123 was chosen as a substrate and different concentrations of hydroxyzine were regard used as inhibitors to assess the role of Pgp in drug interaction.
The result showed that hydroxyzine did not increase the accumulation of R123 in MDR-transfected HEK293 cells. Besides of R site, there is still the other binding site (H site) on Pgp. For this reason, colchicine, the substrate of H site, was used to investigate the interaction with hydroxyzine on Pgp. Similar to R123, hydroxyzine did not increase the accumulation of colchicine in MDR-transfected HEK293 cells. According to above results, we cannot prove that hydroxyzine is able to increase the accumulation of Pgp substrates.
目錄
誌謝 1
中文摘要 2
英文摘要 3
縮寫表 4
目錄 5
表目錄 7
圖目錄 8
第一章. 緒論 10
第一節. 藥物簡介 10
第二節. 多重抗藥性 11
第三節. 多重抗藥性的種類及Pgp的表現 11
第四節. Pgp的結構、功能及其分佈 12
第五節. Pgp在藥物交互作用的角色 13
第六節. Pgp的受質、作用機轉及其結合位置 14
第七節. Pgp的抑制劑及其調控機轉 15
第二章. 研究目的 26
第三章. 實驗材料及方法 27
第一節. 材料 27
第二節. 儀器 36
第三節. 實驗方法 37
第四章. 實驗結果 49
第一節. Construction of pcDNA3-hMDR1 49
第二節. Pgp在MDR-transfected HEK細胞中的表現 49
第三節. 細胞毒性試驗 50
第四節. Rhodamine123在HEK及HEK-MDR細胞株中的uptake的情形 51
第五節. [3H]-Colchicine在HEK及HEK-MDR細胞株中的uptake的情形 53
第五章. 討論 70
參考文獻 73
參考文獻
Ambudkar, S.V., Dey, S., Hrycyna, C.A., Ramachandra, M., Pastan, I. and Gottesman, M.M. (1999) Biochemical, cellular, and pharmacological aspects of the multidrug transporter. Annu Rev Pharmacol Toxicol 39, 361-98.
Bech-Hansen, N.T., Till, J.E. and Ling, V. (1976) Pleiotropic phenotype of colchicine-resistant CHO cells: cross- resistance and collateral sensitivity. J Cell Physiol 88, 23-31.
Biedler, J.L. and Riehm, H. (1970) Cellular resistance to actinomycin D in Chinese hamster cells in vitro: cross-resistance, radioautographic, and cytogenetic studies. Cancer Res 30, 1174-84.
Boer, R., Gekeler, V., Ulrich, W.R., Zimmermann, P., Ise, W., Schodl, A. and Haas, S. (1996) Modulation of P-glycoprotein mediated drug accumulation in multidrug resistant CCRF VCR-1000 cells by chemosensitisers. Eur J Cancer 32A, 857-61.
Borst, P. (1997) Multidrug resistant proteins. Semin Cancer Biol 8, 131-4.
Borst, P. and Schinkel, A.H. (1997) Genetic dissection of the function of mammalian P-glycoproteins. Trends Genet 13, 217-22.
Chen, C.J., Chin, J.E., Ueda, K., Clark, D.P., Pastan, I., Gottesman, M.M. and Roninson, I.B. (1986) Internal duplication and homology with bacterial transport proteins in the mdr1 (P-glycoprotein) gene from multidrug-resistant human cells. Cell 47, 381-9.
Chen, C.J., Clark, D., Ueda, K., Pastan, I., Gottesman, M.M. and Roninson, I.B. (1990) Genomic organization of the human multidrug resistance (MDR1) gene and origin of P-glycoproteins. J Biol Chem 265, 506-14.
Cordon-Cardo, C., O'Brien, J.P., Boccia, J., Casals, D., Bertino, J.R. and Melamed, M.R. (1990) Expression of the multidrug resistance gene product (P-glycoprotein) in human normal and tumor tissues. J Histochem Cytochem 38, 1277-87.
Daoud, R., Kast, C., Gros, P. and Georges, E. (2000) Rhodamine 123 binds to multiple sites in the multidrug resistance protein (MRP1). Biochemistry 39, 15344-52.
Fisher, G.A., Lum, B.L., Hausdorff, J. and Sikic, B.I. (1996) Pharmacological considerations in the modulation of multidrug resistance. Eur J Cancer 32A, 1082-8.
Fojo, A., Akiyama, S., Gottesman, M.M. and Pastan, I. (1985) Reduced drug accumulation in multiply drug-resistant human KB carcinoma cell lines. Cancer Res 45, 3002-7.
Ford, J.M. (1995) Modulators of multidrug resistance. Preclinical studies. Hematol Oncol Clin North Am 9, 337-61.
Gengo, F.M., Dabronzo, J., Yurchak, A., Love, S. and Miller, J.K. (1987) The relative antihistaminic and psychomotor effects of hydroxyzine and cetirizine. Clin Pharmacol Ther 42, 265-72.
Germann, U.A., Ford, P.J., Shlyakhter, D., Mason, V.S. and Harding, M.W. (1997) Chemosensitization and drug accumulation effects of VX-710, verapamil, cyclosporin A, MS-209 and GF120918 in multidrug resistant HL60/ADR cells expressing the multidrug resistance-associated protein MRP. Anticancer Drugs 8, 141-55.
Greenberger, L.M. (1993) Major photoaffinity drug labeling sites for iodoaryl azidoprazosin in P- glycoprotein are within, or immediately C-terminal to, transmembrane domains 6 and 12. J Biol Chem 268, 11417-25.
Higgins, C.F. and Gottesma, M.M. (1992) Is the multidrug transporter a flippase? Trends Biochem Sci 17, 18-21.
Hirsch-Ernst, K.I., Ziemann, C., Rustenbeck, I. and Kahl, G.F. (2001) Inhibitors of mdr1-dependent transport activity delay accumulation of the mdr1 substrate rhodamine 123 in primary rat hepatocyte cultures. Toxicology 167, 47-57.
Hori, R., Okamura, N., Aiba, T. and Tanigawara, Y. (1993) Role of P-glycoprotein in renal tubular secretion of digoxin in the isolated perfused rat kidney. J Pharmacol Exp Ther 266, 1620-5.
Hsu, S.I., Lothstein, L. and Horwitz, S.B. (1989) Differential overexpression of three mdr gene family members in multidrug-resistant J774.2 mouse cells. Evidence that distinct P- glycoprotein precursors are encoded by unique mdr genes. J Biol Chem 264, 12053-62.
Hyde, S.C., Emsley, P., Hartshorn, M.J., Mimmack, M.M., Gileadi, U., Pearce, S.R., Gallagher, M.P., Gill, D.R., Hubbard, R.E. and Higgins, C.F. (1990) Structural model of ATP-binding proteins associated with cystic fibrosis, multidrug resistance and bacterial transport. Nature 346, 362-5.
Johnson, L.V., Walsh, M.L. and Chen, L.B. (1980) Localization of mitochondria in living cells with rhodamine 123. Proc Natl Acad Sci U S A 77, 990-4.
Juliano, R.L. and Ling, V. (1976) A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. Biochim Biophys Acta 455, 152-62.
Kan, W.M., Liu, Y.T., Hsiao, C.L., Shieh, C.Y., Kuo, J.H., Huang, J.D. and Su, S.F. (2001) Effect of hydroxyzine on the transport of etoposide in rat small intestine. Anticancer Drugs 12, 267-73.
Kusunoki, N., Takara, K., Tanigawara, Y., Yamauchi, A., Ueda, K., Komada, F., Ku, Y., Kuroda, Y., Saitoh, Y. and Okumura, K. (1998) Inhibitory effects of a cyclosporin derivative, SDZ PSC 833, on transport of doxorubicin and vinblastine via human P-glycoprotein. Jpn J Cancer Res 89, 1220-8.
Lincke, C.R., Smit, J.J., van der Velde-Koerts, T. and Borst, P. (1991) Structure of the human MDR3 gene and physical mapping of the human MDR locus. J Biol Chem 266, 5303-10.
Loo, T.W. and Clarke, D.M. (1994) Reconstitution of drug-stimulated ATPase activity following co- expression of each half of human P-glycoprotein as separate polypeptides. J Biol Chem 269, 7750-5.
Loo, T.W. and Clarke, D.M. (1995) Rapid purification of human P-glycoprotein mutants expressed transiently in HEK 293 cells by nickel-chelate chromatography and characterization of their drug-stimulated ATPase activities. J Biol Chem 270, 21449-52.
Lum, B.L., Fisher, G.A., Brophy, N.A., Yahanda, A.M., Adler, K.M., Kaubisch, S., Halsey, J. and Sikic, B.I. (1993) Clinical trials of modulation of multidrug resistance. Pharmacokinetic and pharmacodynamic considerations. Cancer 72, 3502-14.
Martin, C., Berridge, G., Higgins, C.F., Mistry, P., Charlton, P. and Callaghan, R. (2000) Communication between multiple drug binding sites on P-glycoprotein. Mol Pharmacol 58, 624-32.
Ozols, R.F., Cunnion, R.E., Klecker, R.W., Jr., Hamilton, T.C., Ostchega, Y., Parrillo, J.E. and Young, R.C. (1987) Verapamil and adriamycin in the treatment of drug-resistant ovarian cancer patients. J Clin Oncol 5, 641-7.
Paton, D.M. and Webster, D.R. (1985) Clinical pharmacokinetics of H1-receptor antagonists (the antihistamines). Clin Pharmacokinet 10, 477-97.
Pawagi, A.B., Wang, J., Silverman, M., Reithmeier, R.A. and Deber, C.M. (1994) Transmembrane aromatic amino acid distribution in P-glycoprotein. A functional role in broad substrate specificity. J Mol Biol 235, 554-64.
Rao, V.V., Dahlheimer, J.L., Bardgett, M.E., Snyder, A.Z., Finch, R.A., Sartorelli, A.C. and Piwnica-Worms, D. (1999) Choroid plexus epithelial expression of MDR1 P glycoprotein and multidrug resistance-associated protein contribute to the blood- cerebrospinal-fluid drug-permeability barrier. Proc Natl Acad Sci U S A 96, 3900-5.
Robert, J. (1999) Multidrug resistance in oncology: diagnostic and therapeutic approaches. Eur J Clin Invest 29, 536-45.
Sand, A.E., Ostlund, E., Andersson, E. and Fried, G. (1998) Endothelin-induced contractions in placental arteries is mediated by both ETA- and ETB-receptors. Acta Physiol Scand 163, 227-34.
Schinkel, A.H., Smit, J.J., van Tellingen, O., Beijnen, J.H., Wagenaar, E., van Deemter, L., Mol, C.A., van der Valk, M.A., Robanus-Maandag, E.C., te Riele, H.P. and et al. (1994) Disruption of the mouse mdr1a P-glycoprotein gene leads to a deficiency in the
blood-brain barrier and to increased sensitivity to drugs. Cell 77, 491-502.
Schinkel, A.H., Wagenaar, E., Mol, C.A. and van Deemter, L. (1996) P-glycoprotein in the blood-brain barrier of mice influences the brain penetration and pharmacological activity of many drugs. J Clin Invest 97, 2517-24.
Shapiro, A.B., Corder, A.B. and Ling, V. (1997) P-glycoprotein-mediated Hoechst 33342 transport out of the lipid bilayer. Eur J Biochem 250, 115-21.
Shapiro, A.B., Fox, K., Lam, P. and Ling, V. (1999) Stimulation of P-glycoprotein-mediated drug transport by prazosin and progesterone. Evidence for a third drug-binding site. Eur J Biochem 259, 841-50.
Shapiro, A.B. and Ling, V. (1997) Positively cooperative sites for drug transport by P-glycoprotein with distinct drug specificities. Eur J Biochem 250, 130-7.
Simons, F.E. (1990) Recent advances in H1-receptor antagonist treatment. J Allergy Clin Immunol 86, 995-9.
Simons, F.E. and Simons, K.J. (1991) Pharmacokinetic optimisation of histamine H1-receptor antagonist therapy. Clin Pharmacokinet 21, 372-93.
Simons, F.E., Simons, K.J., Chung, M. and Yeh, J. (1987) The comparative pharmacokinetics of H1-receptor antagonists. Ann Allergy 59, 20-4.
Simons, F.E., Simons, K.J. and Frith, E.M. (1984) The pharmacokinetics and antihistaminic of the H1 receptor antagonist hydroxyzine. J Allergy Clin Immunol 73, 69-75.
Stapf, V., Thalhammer, T., Huber-Huber, R., Felberbauer, F., Gajdzik, L. and Graf, J. (1994) Inhibition of rhodamine 123 secretion by cyclosporin A as a model of P- glycoprotein mediated transport in liver. Anticancer Res 14, 581-5.
Stouch, T.R. and Gudmundsson, O. (2002) Progress in understanding the structure-activity relationships of P- glycoprotein. Adv Drug Deliv Rev 54, 315-28.
Takada, Y., Yamada, K., Taguchi, Y., Kino, K., Matsuo, M., Tucker, S.J., Komano, T., Amachi, T. and Ueda, K. (1998) Non-equivalent cooperation between the two nucleotide-binding folds of P-glycoprotein. Biochim Biophys Acta 1373, 131-6.
Tamai, I. and Safa, A.R. (1991) Azidopine noncompetitively interacts with vinblastine and cyclosporin A binding to P-glycoprotein in multidrug resistant cells. J Biol Chem 266, 16796-800.
Tan, B., Piwnica-Worms, D. and Ratner, L. (2000) Multidrug resistance transporters and modulation. Curr Opin Oncol 12, 450-8.
Tanigawara, Y. (2000) Role of P-glycoprotein in drug disposition. Ther Drug Monit 22, 137-40.
Traunecker, H.C., Stevens, M.C., Kerr, D.J. and Ferry, D.R. (1999) The acridonecarboxamide GF120918 potently reverses P-glycoprotein- mediated resistance in human sarcoma MES-Dx5 cells. Br J Cancer 81, 942-51.
Tsuruo, T., Iida, H., Tsukagoshi, S. and Sakurai, Y. (1981) Overcoming of vincristine resistance in P388 leukemia in vivo and in vitro through enhanced cytotoxicity of vincristine and vinblastine by verapamil. Cancer Res 41, 1967-72.
Ueda, K., Clark, D.P., Chen, C.J., Roninson, I.B., Gottesman, M.M. and Pastan, I. (1987) The human multidrug resistance (mdr1) gene. cDNA cloning and transcription initiation. J Biol Chem 262, 505-8.
Ueda, K., Yoshida, A. and Amachi, T. (1999) Recent progress in P-glycoprotein research. Anticancer Drug Des 14, 115-21.
Urbatsch, I.L., Sankaran, B., Bhagat, S. and Senior, A.E. (1995) Both P-glycoprotein nucleotide-binding sites are catalytically active. J Biol Chem 270, 26956-61.
van der Bliek, A.M., Kooiman, P.M., Schneider, C. and Borst, P. (1988) Sequence of mdr3 cDNA encoding a human P-glycoprotein. Gene 71, 401-11.
Wacher, V.J., Wu, C.Y. and Benet, L.Z. (1995) Overlapping substrate specificities and tissue distribution of cytochrome P450 3A and P-glycoprotein: implications for drug delivery and activity in cancer chemotherapy. Mol Carcinog 13, 129-34.
Wood, S.G., John, B.A., Chasseaud, L.F., Yeh, J. and Chung, M. (1987) The metabolism and pharmacokinetics of 14C-cetirizine in humans. Ann Allergy 59, 31-4.
Zeng, H., Bain, L.J., Belinsky, M.G. and Kruh, G.D. (1999) Expression of multidrug resistance protein-3 (multispecific organic anion transporter-D) in human embryonic kidney 293 cells confers resistance to anticancer agents. Cancer Res 59, 5964-7.
連結至畢業學校之論文網頁點我開啟連結
註: 此連結為研究生畢業學校所提供,不一定有電子全文可供下載,若連結有誤,請點選上方之〝勘誤回報〞功能,我們會盡快修正,謝謝!
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關期刊