請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26436完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 朱善德 | |
| dc.contributor.author | Chuan-Jen Hung | en |
| dc.contributor.author | 洪蓴仁 | zh_TW |
| dc.date.accessioned | 2021-06-08T07:10:12Z | - |
| dc.date.copyright | 2008-08-04 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-07-31 | |
| dc.identifier.citation | Akerstrom B, Flower DR & Salier JP 2000 Lipocalins: unity in diversity. Biochim
Biophys Acta 1482 1-8. Arici A, Head JR, MacDonald PC & Casey ML 1993 Regulation of interleukin-8 gene expression in human endometrial cells in culture. Mol Cell Endocrinol 94 195-204. Arici A, Seli E, Senturk LM, Gutierrez LS, Oral E & Taylor HS 1998a Interleukin-8 in the human endometrium. J Clin Endocrinol Metab 83 1783-1787. Arici A, Seli E, Zeyneloglu HB, Senturk LM, Oral E & Olive DL 1998b Interleukin-8 induces proliferation of endometrial stromal cells: a potential autocrine growth factor. J Clin Endocrinol Metab 83 1201-1205. Arici A, Senturk LM, Seli E, Bahtiyar MO & Kim G 1999 Regulation of monocyte chemotactic protein-1 expression in human endometrial stromal cells by estrogen and progesterone. Biol Reprod 61 85-90. Arici A, Tazuke SI, Attar E, Kliman HJ & Olive DL 1996 Interleukin-8 concentration in peritoneal fluid of patients with endometriosis and modulation of interleukin-8 expression in human mesothelial cells. Mol Hum Reprod 2 40-45. Barcelos LS, Talvani A, Teixeira AS, Cassali GD, Andrade SP & Teixeira MM 2004 Production and in vivo effects of chemokines CXCL1-3/KC and CCL2/JE in a model of inflammatory angiogenesis in mice. Inflamm Res 53 576-584. Beyaert R, Cuenda A, Vanden Berghe W, Plaisance S, Lee JC, Haegeman G, Cohen P & Fiers W 1996 The p38/RK mitogen-activated protein kinase pathway regulates interleukin-6 synthesis response to tumor necrosis factor. EMBO J 15 1914-1923. Bong JJ, Seol MB, Kim HH, Han O, Back K & Baik M 2004 The 24p3 gene is induced during involution of the mammary gland and induces apoptosis of mammary epithelial cells. Mol Cells 17 29-34. 94 Bozic CR, Gerard NP, von Uexkull-Guldenband C, Kolakowski LF, Jr., Conklyn MJ, Breslow R, Showell HJ & Gerard C 1994 The murine interleukin 8 type B receptor homologue and its ligands. Expression and biological characterization. J Biol Chem 269 29355-29358. Bratt T 2000 Lipocalins and cancer. Biochim Biophys Acta 1482 318-326. Brook M, Sully G, Clark AR & Saklatvala J 2000 Regulation of tumour necrosis factor alpha mRNA stability by the mitogen-activated protein kinase p38 signalling cascade. FEBS Lett 483 57-61. Caivano M & Cohen P 2000 Role of mitogen-activated protein kinase cascades in mediating lipopolysaccharide-stimulated induction of cyclooxygenase-2 and IL-1 beta in RAW264 macrophages. J Immunol 164 3018-3025. Chen CC & Wang JK 1999 p38 but not p44/42 mitogen-activated protein kinase is required for nitric oxide synthase induction mediated by lipopolysaccharide in RAW 264.7 macrophages. Mol Pharmacol 55 481-488. Coles M, Diercks T, Muehlenweg B, Bartsch S, Zolzer V, Tschesche H & Kessler H 1999 The solution structure and dynamics of human neutrophil gelatinase-associated lipocalin. J Mol Biol 289 139-157. Cui S, Reichner JS, Mateo RB & Albina JE 1994 Activated murine macrophages induce apoptosis in tumor cells through nitric oxide-dependent or -independent mechanisms. Cancer Res 54 2462-2467. Davis TR, Tabatabai L, Bruns K, Hamilton RT & Nilsen-Hamilton M 1991 Basic fibroblast growth factor induces 3T3 fibroblasts to synthesize and secrete a cyclophilin-like protein and beta 2-microglobulin. Biochim Biophys Acta 1095 145-152. Devireddy LR, Gazin C, Zhu X & Green MR 2005 A cell-surface receptor for lipocalin 24p3 selectively mediates apoptosis and iron uptake. Cell 123 1293-1305. Devireddy LR, Teodoro JG, Richard FA & Green MR 2001 Induction of apoptosis by a secreted lipocalin that is transcriptionally regulated by IL-3 deprivation. Science 293 829-834. 95 Driscoll KE, Hassenbein DG, Howard BW, Isfort RJ, Cody D, Tindal MH, Suchanek M & Carter JM 1995 Cloning, expression, and functional characterization of rat MIP-2: a neutrophil chemoattractant and epithelial cell mitogen. J Leukoc Biol 58 359-364. Elangovan N, Lee YC, Tzeng WF & Chu ST 2004 Delivery of ferric ion to mouse spermatozoa is mediated by lipocalin internalization. Biochem Biophys Res Commun 319 1096-1104. Fan X, Patera AC, Pong-Kennedy A, Deno G, Gonsiorek W, Manfra DJ, Vassileva G, Zeng M, Jackson C, Sullivan L, et al. 2007 Murine CXCR1 is a functional receptor for GCP-2/CXCL6 and interleukin-8/CXCL8. J Biol Chem 282 11658-11666. Farias-Eisner R, Sherman MP, Aeberhard E & Chaudhuri G 1994 Nitric oxide is an important mediator for tumoricidal activity in vivo. Proc Natl Acad Sci U S A 91 9407-9411. Finn CA & Publicover M 1981 Hormonal control of cell death in the luminal epithelium of the mouse uterus. J Endocrinol 91 335-340. Flower DR 1996 The lipocalin protein family: structure and function. Biochem J 318 ( Pt 1) 1-14. Garcia-Velasco JA & Arici A 1999 Chemokines and human reproduction. Fertil Steril 71 983-993. Gijsbers K, Gouwy M, Struyf S, Wuyts A, Proost P, Opdenakker G, Penninckx F, Ectors N, Geboes K & Van Damme J 2005 GCP-2/CXCL6 synergizes with other endothelial cell-derived chemokines in neutrophil mobilization and is associated with angiogenesis in gastrointestinal tumors. Exp Cell Res 303 331-342. Goodman RB, Wood RG, Martin TR, Hanson-Painton O & Kinasewitz GT 1992 Cytokine-stimulated human mesothelial cells produce chemotactic activity for neutrophils including NAP-1/IL-8. J Immunol 148 457-465. Greb RR, Heikinheimo O, Williams RF, Hodgen GD & Goodman AL 1997 Vascular endothelial growth factor in primate endometrium is regulated by oestrogen-receptor and progesterone-receptor ligands in vivo. Hum Reprod 12 1280-1292. 96 Hraba-Renevey S, Turler H, Kress M, Salomon C & Weil R 1989 SV40-induced expression of mouse gene 24p3 involves a post-transcriptional mechanism. Oncogene 4 601-608. Huang HL, Chu ST & Chen YH 1999 Ovarian steroids regulate 24p3 expression in mouse uterus during the natural estrous cycle and the preimplantation period. J Endocrinol 162 11-19. Huang J, Roby KF, Pace JL, Russell SW & Hunt JS 1995 Cellular localization and hormonal regulation of inducible nitric oxide synthase in cycling mouse uterus. J Leukoc Biol 57 27-35. Huang Y, Zhu XY, Du MR, Wu X, Wang MY & Li DJ 2006 Chemokine CXCL16, a scavenger receptor, induces proliferation and invasion of first-trimester human trophoblast cells in an autocrine manner. Hum Reprod 21 1083-1091. Karin M & Hunter T 1995 Transcriptional control by protein phosphorylation: signal transmission from the cell surface to the nucleus. Curr Biol 5 747-757. Kayisli UA, Mahutte NG & Arici A 2002 Uterine chemokines in reproductive physiology and pathology. Am J Reprod Immunol 47 213-221. Kim DS, Han JH & Kwon HJ 2003 NF-kappaB and c-Jun-dependent regulation of macrophage inflammatory protein-2 gene expression in response to lipopolysaccharide in RAW 264.7 cells. Mol Immunol 40 633-643. Kjeldsen L, Cowland JB & Borregaard N 2000 Human neutrophil gelatinase-associated lipocalin and homologous proteins in rat and mouse. Biochim Biophys Acta 1482 272-283. Koch AE, Polverini PJ, Kunkel SL, Harlow LA, DiPietro LA, Elner VM, Elner SG & Strieter RM 1992 Interleukin-8 as a macrophage-derived mediator of angiogenesis. Science 258 1798-1801. Kollmar O, Scheuer C, Menger MD & Schilling MK 2006 Macrophage inflammatory protein-2 promotes angiogenesis, cell migration, and tumor growth in hepatic metastasis. Ann Surg Oncol 13 263-275. 97 Kreuz S, Siegmund D, Rumpf JJ, Samel D, Leverkus M, Janssen O, Hacker G, Dittrich-Breiholz O, Kracht M, Scheurich P, et al. 2004 NFkappaB activation by Fas is mediated through FADD, caspase-8, and RIP and is inhibited by FLIP. J Cell Biol 166 369-380. Lai MD, Lee LR, Cheng KS & Wing LY 2000 Expression of proliferating cell nuclear antigen in luminal epithelium during the growth and regression of rat uterus. J Endocrinol 166 87-93. Larsen CG, Anderson AO, Oppenheim JJ & Matsushima K 1989 Production of interleukin-8 by human dermal fibroblasts and keratinocytes in response to interleukin-1 or tumour necrosis factor. Immunology 68 31-36. Lin HH, Li WW, Lee YC & Chu ST 2007 Apoptosis induced by uterine 24p3 protein in endometrial carcinoma cell line. Toxicology 234 203-215. Litwin S, Lagadari M, Barrientos G, Roux ME, Margni R & Miranda S 2005 Comparative immunohistochemical study of M-CSF and G-CSF in feto-maternal interface in a multiparity mouse model. Am J Reprod Immunol 54 311-320. Liu Q & Nilsen-Hamilton M 1995 Identification of a new acute phase protein. J Biol Chem 270 22565-22570. Logdberg L & Wester L 2000 Immunocalins: a lipocalin subfamily that modulates immune and inflammatory responses. Biochim Biophys Acta 1482 284-297. Lorsbach RB, Murphy WJ, Lowenstein CJ, Snyder SH & Russell SW 1993 Expression of the nitric oxide synthase gene in mouse macrophages activated for tumor cell killing. Molecular basis for the synergy between interferon-gamma and lipopolysaccharide. J Biol Chem 268 1908-1913. Ma W, Miao Z & Novotny MV 1998 Role of the adrenal gland and adrenal-mediated chemosignals in suppression of estrus in the house mouse: the lee-boot effect revisited. Biol Reprod 59 1317-1320. Ma W, Miao Z & Novotny MV 1999 Induction of estrus in grouped female mice (Mus domesticus) by synthetic analogues of preputial gland constituents. Chem Senses 24 289-293. 98 Martin L, Pollard JW & Fagg B 1976 Oestriol, oestradiol-17beta and the proliferation and death of uterine cells. J Endocrinol 69 103-115. Meheus LA, Fransen LM, Raymackers JG, Blockx HA, Van Beeumen JJ, Van Bun SM & Van de Voorde A 1993 Identification by microsequencing of lipopolysaccharide-induced proteins secreted by mouse macrophages. J Immunol 151 1535-1547. Pollard JW, Lin EY & Zhu L 1998 Complexity in uterine macrophage responses to cytokines in mice. Biol Reprod 58 1469-1475. Ren X, Carpenter A, Hogaboam C & Colletti L 2003 Mitogenic properties of endogenous and pharmacological doses of macrophage inflammatory protein-2 after 70 % hepatectomy in the mouse. Am J Pathol 163 563-570. Rollins BJ 1997 Chemokines. Blood 90 909-928. Rovai LE, Herschman HR & Smith JB 1998 The murine neutrophil-chemoattractant chemokines LIX, KC, and MIP-2 have distinct induction kinetics, tissue distributions, and tissue-specific sensitivities to glucocorticoid regulation in endotoxemia. J Leukoc Biol 64 494-502. Ryan IP, Tseng JF, Schriock ED, Khorram O, Landers DV & Taylor RN 1995 Interleukin-8 concentrations are elevated in peritoneal fluid of women with endometriosis. Fertil Steril 63 929-932. Ryon J, Bendickson L & Nilsen-Hamilton M 2002 High expression in involuting reproductive tissues of uterocalin/24p3, a lipocalin and acute phase protein. Biochem J 367 271-277. Sato T, Fukazawa Y, Kojima H, Enari M, Iguchi T & Ohta Y 1997 Apoptotic cell death during the estrous cycle in the rat uterus and vagina. Anat Rec 248 76-83. Schadendorf D, Moller A, Algermissen B, Worm M, Sticherling M & Czarnetzki BM 1993 IL-8 produced by human malignant melanoma cells in vitro is an essential autocrine growth factor. J Immunol 151 2667-2675. 99 Schmidt HH, Warner TD, Nakane M, Forstermann U & Murad F 1992 Regulation and subcellular location of nitrogen oxide synthases in RAW264.7 macrophages. Mol Pharmacol 41 615-624. Shifren JL, Tseng JF, Zaloudek CJ, Ryan IP, Meng YG, Ferrara N, Jaffe RB & Taylor RN 1996 Ovarian steroid regulation of vascular endothelial growth factor in the human endometrium: implications for angiogenesis during the menstrual cycle and in the pathogenesis of endometriosis. J Clin Endocrinol Metab 81 3112-3118. Strieter RM, Kunkel SL, Showell HJ, Remick DG, Phan SH, Ward PA & Marks RM 1989 Endothelial cell gene expression of a neutrophil chemotactic factor by TNF-alpha, LPS, and IL-1 beta. Science 243 1467-1469. Strieter RM, Polverini PJ, Arenberg DA, Walz A, Opdenakker G, Van Damme J & Kunkel SL 1995a Role of C-X-C chemokines as regulators of angiogenesis in lung cancer. J Leukoc Biol 57 752-762. Strieter RM, Polverini PJ, Kunkel SL, Arenberg DA, Burdick MD, Kasper J, Dzuiba J, Van Damme J, Walz A, Marriott D, et al. 1995b The functional role of the ELR motif in CXC chemokine-mediated angiogenesis. J Biol Chem 270 27348-27357. Stuehr DJ & Griffith OW 1992 Mammalian nitric oxide synthases. Adv Enzymol Relat Areas Mol Biol 65 287-346. Tibbetts TA, Conneely OM & O'Malley BW 1999 Progesterone via its receptor antagonizes the pro-inflammatory activity of estrogen in the mouse uterus. Biol Reprod 60 1158-1165. Tong Z, Wu X, Ovcharenko D, Zhu J, Chen CS & Kehrer JP 2005 Neutrophil gelatinase-associated lipocalin as a survival factor. Biochem J 391 441-448. Tuschil A, Lam C, Haslberger A & Lindley I 1992 Interleukin-8 stimulates calcium transients and promotes epidermal cell proliferation. J Invest Dermatol 99 294-298. Van Coillie E, Van Aelst I, Wuyts A, Vercauteren R, Devos R, De Wolf-Peeters C, Van Damme J & Opdenakker G 2001 Tumor angiogenesis induced by granulocyte chemotactic protein-2 as a countercurrent principle. Am J Pathol 159 1405-1414. 100 Walmer DK, Wrona MA, Hughes CL & Nelson KG 1992 Lactoferrin expression in the mouse reproductive tract during the natural estrous cycle: correlation with circulating estradiol and progesterone. Endocrinology 131 1458-1466. Wu R, Van der Hoek KH, Ryan NK, Norman RJ & Robker RL 2004 Macrophage contributions to ovarian function. Hum Reprod Update 10 119-133. Wuyts A, D'Haese A, Cremers V, Menten P, Lenaerts JP, De Loof A, Heremans H, Proost P & Van Damme J 1999 NH2- and COOH-terminal truncations of murine granulocyte chemotactic protein-2 augment the in vitro and in vivo neutrophil chemotactic potency. J Immunol 163 6155-6163. Wuyts A, Haelens A, Proost P, Lenaerts JP, Conings R, Opdenakker G & Van Damme J 1996 Identification of mouse granulocyte chemotactic protein-2 from fibroblasts and epithelial cells. Functional comparison with natural KC and macrophage inflammatory protein-2. J Immunol 157 1736-1743. Yin Y, Huang WW, Lin C, Chen H, MacKenzie A & Ma L 2008 Estrogen suppresses uterine epithelial apoptosis by inducing birc1 expression. Mol Endocrinol 22 113-125. Yue TL, Wang X, Sung CP, Olson B, McKenna PJ, Gu JL & Feuerstein GZ 1994 Interleukin-8. A mitogen and chemoattractant for vascular smooth muscle cells. Circ Res 75 1-7. Zhang JS, Feng WG, Li CL, Wang XY & Chang ZL 2000 NF-kappa B regulates the LPS-induced expression of interleukin 12 p40 in murine peritoneal macrophages: roles of PKC, PKA, ERK, p38 MAPK, and proteasome. Cell Immunol 204 38-45. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26436 | - |
| dc.description.abstract | 24p3 蛋白是小鼠子宮液中之分泌性蛋白，研究顯示24p3 蛋白的表現受到動情素
與黃體素之調節，主要表現於動情前期與動情期的子宮組織，於動情後期與動情 間期表現量很少，暗示24p3 蛋白於雌性生殖系統扮演重要的角色。利用人類子宮 內膜細胞株RL95-2 探討24p3 蛋白於子宮的生理功能，24p3 蛋白會造成子宮內膜 細胞caspase-3 活性增加並伴隨細胞凋亡，長時間處理下24p3 蛋白會誘發IL-8 的 產生並抑制caspase-3 活性而促使細胞存活，此現象可能與動情週期子宮組織重整 有關。因此本論文利用小鼠動情週期探討24p3 蛋白與IL-8 於子宮組織的相關表現 以了解24p3 蛋白與子宮組織重整的關係。 以西方點墨法、免疫組織化學染色法以及原位末端標記法確認動情期與動情後 期子宮組織有細胞凋亡的現象。於動情週期中24p3 mRNA 與三個IL-8 功能性同源 蛋白 (MIP-2、KC 以及GCP-2) 的mRNA 皆於動情期有顯著的表現，24p3 蛋白可 能藉由誘發小鼠IL-8 功能性同源蛋白而抑制24p3 蛋白引發的子宮內膜細胞凋亡， 參與子宮組織重整的現象。由於巨噬細胞大量存在於動情期子宮組織並為細胞激 素重要來源，實驗中發現24p3 蛋白可活化小鼠巨噬細胞株RAW 264.7 促進一氧化 氮 (NO) 的釋放，此一活化現象可能透過iNOS 以及TNF-α 基因表現量上升進而 促使巨噬細胞株凋亡。24p3 蛋白活化巨噬細胞亦會誘導MIP-2 與KC 的表現，對 GCP-2 表現則無影響，此結果與LPS 活化巨噬細胞的結果相同。根據實驗數據推 測24p3 蛋白可能藉由ERK 與JNK 來影響巨噬細胞相關基因之表現。同時本論文 亦初步建立小鼠子宮內膜上皮細胞初級培養系統，以利研究24p3 蛋白與小鼠子宮 內膜細胞凋亡以及相關細胞激素分泌之關係，期望提供較多資訊以瞭解24p3 蛋白 於生殖系統中可能的生理功能。 | zh_TW |
| dc.description.abstract | 24p3 protein has been implicated in the cell death. Supplementation of 24p3 protein
in human endometrial cell line (RL95-2) causes the elevation of intracellular ROS and activation of caspases, resulting in cell apoptosis. In addition, 24p3 protein induced-cell apoptosis could be suppressed by persistent incubation with 24p3 protein via cytokine secretion. It implied 24p3 protein involved in the uterine remodeling during the physiological cycle. In order to demonstrate such hypothesize we attempt to investigate the expression of 24p3 protein, IL-8 and apoptotic index in the uterus throughout the mouse estrous cycle. 24p3 protein secreted in proestrus and estrus abundantly, and then declined from metestrus to diestrus. Using Western analyses, IHC and TUNEL to measure apoptotic index of endometrial cells during the mouse estrous cycle, we found that 24p3 protein expression coincided with the time scale of endometrial cell apoptosis during estrous cycle. It suggests 24p3 protein play a role on the viability of endometrial cells directly. Furthermore, the expression of 24p3 mRNA ,and three murine functional IL-8 homologues (MIP-2, KC,and GCP-2) mRNA in uterus were much higher in estrous stage during the estrous cycle. 24p3 protein may upregulate the expression of these murine cytokines and inhibit 24p3 protein-induced apoptosis. Because of macrophages are the major source of the cytokines in biological system and are abundant in the estrous stage, we have paid more attention on evaluating the influence of 24p3 protein on murine macrophage. Using macrophage cell line (RAW 264.7) , we found that 24p3 protein can trigger the nitric oxide releasing in a time- and dose-dependent manner on the cells via iNOS and TNF-α gene expression. Besides, 24p3 protein can induce macrophage activation by increasing iNOS and TNF-α gene to undergo vi activation-induced cell death. After activation, the gene expressions of MIP-2 and KC but not of GCP-2 are induced by 24p3 protein in macrophages and the results are similar to LPS-activated macrophages. In the meantime, we have established the mouse uterine epithelial primary cell culture in order to extend our goal for elucidating the effect of 24p3 protein on endometrial epithelial cells. This study may provide the information regarding the pathogenesis of uterine disease. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T07:10:12Z (GMT). No. of bitstreams: 1 ntu-97-R95b46023-1.pdf: 3843627 bytes, checksum: 5b2d3f0b614e4b7ad4158327f524583d (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | 目錄 .................................................................................................................................. i
中文摘要 ......................................................................................................................... iv ABSTRACT ........................................................................................................................ v 縮寫表 ............................................................................................................................ vii 第一章 緒論 .................................................................................................................... 1 一、疏水性分子結合蛋白家族 .................................................................................. 1 二、24p3 蛋白 ............................................................................................................. 2 三、小鼠動情週期 ...................................................................................................... 7 四、子宮細胞激素 ...................................................................................................... 8 五、巨噬細胞在子宮扮演的角色 ............................................................................ 10 六、巨噬細胞的活化 ................................................................................................ 12 七、本論文的研究目的 ............................................................................................ 13 第二章 材料與方法 ...................................................................................................... 16 PART 1 小鼠動物實驗 ............................................................................................... 16 一、小鼠子宮液中24p3 蛋白純化 ...................................................................... 16 1.1 子宮液的誘發與存取 .................................................................................. 16 1.2 分子篩膠體管柱層析 .................................................................................. 16 1.3 蛋白質溶液的濃縮 ...................................................................................... 17 1.4 逆向高效能液相層析 .................................................................................. 17 二、小鼠動情週期各時期的判定 ........................................................................ 19 2.1 動情週期同步化 .......................................................................................... 19 2.2 動情週期的判定 .......................................................................................... 19 三、子宮組織 RNA 的分析 ................................................................................. 20 3.1 組織總核醣核酸的萃取 .............................................................................. 20 3.2 反轉錄反應 .................................................................................................. 21 3.3 聚合酶連鎖反應 .......................................................................................... 22 3.4 洋菜膠體電泳 .............................................................................................. 24 四、子宮組織蛋白的萃取 .................................................................................... 24 五、BCA 蛋白質定量 ........................................................................................... 25 六、十二酯硫酸鈉-聚丙烯醯胺膠體電泳 ........................................................... 26 七、西方點墨法 .................................................................................................... 27 八、免疫組織化學染色 ........................................................................................ 29 8.1 組織包埋與切片製作 ................................................................................. 29 ii 8.2 免疫組織化學染色 ...................................................................................... 31 九、原位末端雜交法 ............................................................................................ 33 十、三明治免疫法染細胞激素抗體陣列膜 ........................................................ 34 十一、小鼠子宮內膜上皮細胞初級培養 ............................................................ 35 十二、免疫螢光染色 ............................................................................................ 36 PART 2 小鼠巨噬細胞株實驗 ................................................................................... 37 十三、小鼠巨噬細胞株RAW 264.7 (ATCC TIB-71) 細胞培養 ........................ 37 13.1 細胞培養 .................................................................................................... 37 13.2 細胞冷凍 .................................................................................................... 38 13.3 細胞解凍 .................................................................................................... 39 十四、細胞RNA 的萃取 ...................................................................................... 39 十五、即時定量聚合酶連鎖反應 ........................................................................ 39 十六、一氧化氮的測量 ........................................................................................ 40 十七、 細胞蛋白質的萃取 .................................................................................. 42 十八、Caspase-3 活性偵測 (PhiPhiLux/ CaspaLux kit) ..................................... 43 第三章 結果 .................................................................................................................. 44 一、小鼠動情週期的判定 ........................................................................................ 44 二、小鼠動情週期子宮內膜細胞的凋亡 ................................................................ 44 三、子宮組織中相關基因於動情週期表現之變化 ................................................ 46 四、表現於小鼠動情期子宮液的細胞激素 ............................................................ 47 五、小鼠巨噬細胞株RAW 264.7 具有24p3 蛋白受體 ......................................... 48 六、24p3 蛋白活化小鼠巨噬細胞株RAW 264.7 ................................................... 49 六、24p3 蛋白引發小鼠巨噬細胞株RAW 264.7 細胞凋亡現象 .......................... 51 七、24p3 蛋白誘導小鼠巨噬細胞株RAW 264.7 細胞激素的分泌 ...................... 51 八、24p3 蛋白影響小鼠巨噬細胞MAPK 磷酸化 .................................................. 52 九、小鼠子宮內膜上皮細胞初級培養系統的建立 ................................................ 53 圖次 ............................................................................................................................... 54 圖一、小鼠24p3 蛋白的純化 .................................................................................. 55 圖二、小鼠動情週期不同時期的陰道抹片 ............................................................ 57 圖三、活化態caspase-3 在小鼠動情週期各時期子宮組織蛋白的表現 ............... 59 圖四、免疫組織化學染色法偵測活化態caspase-3 在子宮組織的表現位置 ....... 61 圖五、TUNEL 法偵測子宮組織凋亡細胞 .............................................................. 63 圖六、分析小鼠子宮組織24p3、24p3 蛋白受體、MIP-2、KC、GCP-2 、IL-6 以 及MCP-1 mRNA 於動情週期不同時期的變化 .......................................... 65 圖七、以細胞激素抗體陣列膜偵測動情期子宮液的細胞激素 ............................ 67 圖九、以RT-PCR 確認小鼠巨噬細胞株RAW 264.7 具有24p3 蛋白受體基因 . 71 iii 圖十、24p3 蛋白以及LPS 對小鼠巨噬細胞株產生一氧化氮的影響 .................. 73 圖十一、24p3 蛋白以及LPS 處理小鼠巨噬細胞株對iNOS mRNA 表現量的影響 ................................................................................................................................... 75 圖十二、24p3 蛋白與LPS 處理小鼠巨噬細胞株對TNF-α mRNA 表現量的影響 ................................................................................................................................... 77 圖十三、24p3 蛋白處理誘發小鼠巨噬細胞株細胞凋亡的現象 ........................... 79 圖十四、24p3 蛋白與LPS 處理小鼠巨噬細胞株對MIP-2、KC 與GCP-2 mRNA 表現量的影響 ............................................................................................ 81 圖十五、24p3 蛋白對小鼠巨噬細胞株MAPK 活性於十二小時內之影響 .......... 83 圖十六、小鼠子宮內膜上皮細胞初級培養之純度鑑定 ........................................ 85 第四章 討論 .................................................................................................................. 86 第五章 參考文獻 .......................................................................................................... 93 | |
| dc.language.iso | zh-TW | |
| dc.subject | 24p3蛋白 | zh_TW |
| dc.subject | 細胞激素 | zh_TW |
| dc.subject | 巨噬細胞 | zh_TW |
| dc.subject | cytokines | en |
| dc.subject | 24p3 protein | en |
| dc.subject | macrophage | en |
| dc.title | 小白鼠子宮24p3蛋白與細胞激素之相關性研究 | zh_TW |
| dc.title | Characterization of the relationship between mouse uterine
24p3 protein and cytokines | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 曾婉芳,李明亭,張?仁,黃彥華 | |
| dc.subject.keyword | 24p3蛋白,細胞激素,巨噬細胞, | zh_TW |
| dc.subject.keyword | 24p3 protein,cytokines,macrophage, | en |
| dc.relation.page | 100 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2008-08-01 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科學研究所 | zh_TW |
| 顯示於系所單位： | 生化科學研究所 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-97-1.pdf 未授權公開取用 | 3.75 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。