探討化學治療後,餵食冬蟲夏草對小鼠免疫機制的影響

簡易檢索 / 詳目顯示

回結果列表

研究生：	莊崴琳 Wei-Lin Chuang
論文名稱：	探討化學治療後,餵食冬蟲夏草對小鼠免疫機制的影響 Inflounce of cordyceps sinensis on mouse immune function after chemo-therapy
指導教授：	江啟勳
口試委員:
學位類別：	碩士 Master
系所名稱：	原子科學院 - 生醫工程與環境科學系 Department of Biomedical Engineering and Environmental Sciences
論文出版年：	2005
畢業學年度：	93
語文別：	中文
論文頁數：	57
中文關鍵詞：	冬蟲夏草、免疫機制
外文關鍵詞：	CTL assay, cordyceps sinensis, ELISPOT
相關次數：	點閱：3 下載：0
分享至:	分享至facebook 分享至twitter

查詢本校圖書館目錄查詢臺灣博碩士論文知識加值系統勘誤回報

在傳統的中國社會中，除了西醫療法外，常常會自行服用中草藥來茲補養生，以增加身體的抵抗力及提升生理機能。這種茲補養生的概念類似於西醫中利用細胞激素來提升人體免疫力的想法。本論文中，我們建立了以傳統中草藥經由口服的投藥方式，再以餵食管餵食小鼠冬蟲夏草的模式，來探討化學治療後，冬蟲夏草對於免疫機制的影響。在小鼠皮下植入攝護腺腫瘤細胞(TRAMP C1)，模擬癌症產生，再施予化學藥物：紫杉醇治療後，比較有無餵食冬蟲夏草對腫瘤生長及免疫機能的影響。結果發現餵食冬蟲夏草可以延遲腫瘤生長速率回升的時間，在免疫機能方面，餵食冬蟲夏草可以加速因化學治療減少的白血球恢復、增強T細胞對抗原的敏感度、增進T細胞的毒殺能力，並且能夠使幫助型T細胞活化，分泌IFN-γ。實驗結果顯示，冬蟲夏草具有調控免疫機能的功效，能加速化學治療後免疫機能的恢復，並有效延緩腫瘤復發的時間。

Traditional Chinese Medicine has used Tonics as a drug to tonify the deficient of the body, enhance the resistance of disease, and invigorate the physiological function of the body. The use of Tonics drug is somehow sharing the similar idea of using cytokines to boots up host immunity in Western Medicine. Cordyceps sinensis (CS) has been used to treat many illnesses in Chinese society for a long time. We have studied the influences on mice immune function under the model of feeding CS as an immune adjuvant. Prostate cancer cell lines, TRAMP C1, were injected subcutaneously into the right flank of C57BL/6J mice to establish a tumor model. Compare the effect of tumor growth and immune activity weather feeding CS or not after treated with chemo-drug, Taxol. The results show that the mice treated with CS have delayed tumor re-growth. In immune function assay, uptake of CS recovered the polymorphonuclrar leukocyte level in blood which has decreased by Taxol treatment. It also promoted T cell sensitivity to antigen, increased cytotoxic T cell responses, and activated T cell to secrete interferon -γ . We demonstrate that cordyceps sinensis could modulate the immune response and facilitate the recovery of immune function after chemo-therapy. It also delays tumor re-growth significantly.

英文摘要.............................................Ⅰ
中文摘要.............................................Ⅱ
誌謝 .................................................Ⅲ
目錄 .................................................Ⅳ
第一章 序論 ......................................... 1
1.    攝護腺癌 ...................................... 1
1.1    　背景 .................................... 1
1.2    　攝護腺癌的症狀 .........................1
1.3    　攝護腺癌的診斷 .........................1
1.4    　攝護腺癌的治療 .........................2
1.5    　小鼠的攝護腺腫瘤細胞 .................. 4
2.    冬蟲夏草 ...................................... 5
2.1  冬蟲夏草的歷史.......................... 5
2.2  冬蟲夏草的地位及分類................... 6
2.3  冬蟲夏草的生活史........................ 6
2.4  冬蟲夏草的組成.......................... 6
2.5  冬蟲夏草的療效.......................... 7
3.    紫杉醇...........................................8
       1.3.1  紫杉醇的歷史.............................8
3.2  紫杉醇的作用機轉........................ 8
3.3  紫杉醇的抗癌效果........................ 9
       1.3.4  紫杉醇的副作用......................... 10
4.    實驗目的....................................... 10
第二章    材料與方法.................................... 11
1.    細胞培養....................................... 11
1.1  配置DMEM細胞培養液.................. 11
1.2  各種細胞培養液配方..................... 11
1.3  純化腹腔巨噬細胞....................... 13
1.4  純化脾臟T細胞......................... 14
1.5  細胞繼代.................................15
2.    冬蟲夏草的製備...... .......................... 15
2.1  冬蟲夏草的來源.......................... 15
2.2  冬蟲夏草的萃取.......................... 16
3.    動物實驗........................................16
3.1      動物來源................................ 16
3.2      動物分組................................ 16
3.3      植入TRAMP C1細胞.....................17
3.4      注射紫杉醇.............................. 17
3.5      餵食冬蟲夏草............................17
3.6      腫瘤重量測量............................18
4.    免疫機能測試................................... 18
4.1      血液樣本................................ 18
4.2      劉氏染色法.............................. 18
4.3      細胞毒殺測試............................ 19
4.4      異種混合T細胞培養反應.................20
4.5      XTT assay................................21
4.6      免疫斑點法(ELISPOT) .................. 21
4.7      吞噬能力測試............................ 22
第三章    實驗結果....................................... 23
1.    腫瘤生長曲線................................... 23
2.    血液樣本測試................................... 24
2.1      計算血液細胞總數....................... 24
2.2      劉氏染色法.............................. 24
2.3      血液中多型核白血球總數................ 25
2.4      血液中淋巴球總數....................... 25
3.    T細胞免疫反應測試.............................26
3.1      純化T細胞.............................. 26
3.2      異種混合T細胞反應..................... 27
3.3      檢視T細胞表現INF-γ的情形.............27
3.4      T細胞毒殺能力檢測......................28
4.    腹腔巨噬細胞免疫反應測試..................... 29
4.1      純化腹腔巨噬細胞....................... 30
4.2      測試巨噬細胞吞噬能力..................  30
第四章    討論.............................. .............31
圖表說明.............................. ................37
圖表.............................. .................... 42
參考文獻.............................. ................54

                                

1. Espinoza-Delgado, I. Cancer vaccines. Oncologist, 7 Suppl 3: 20-33, 2002.
2. Smith DC, Dunn RL, Strawderman MS, Pienta KJ. Change in serum prostate-specific antigen as a marker of response to cytotoxic therapy for hormone-refractory prostate cancer. J Clin Oncol., 16: 1835-1843, 1998.
3. 陳明冬蟲夏草中核酸及鹼基測定方法的研究. 中成藥, 17: 40-41, 1995.
4. Yoo HS, Shin JW, Cho JH, Son CG, Lee YW, Park SY, Cho CK. Effects of Cordyceps militaris extract on angiogenesis and tumor growth. Acta Pharmacol Sin., 25: 657~665, 2004.
5. Chen YJ, Shiao MS, Lee SS, Wang SY. Effect of Cordyceps sinensis on the proliferation and differentiation of human leukemic U937 cells. Life Sci., 60: 2349~2359, 1997.
6. Yoshida J, Takamura S, Yamaguchi N, et al. Antitumor activity of an extract of Cordyceps sinensis (Berk.) Sacc. against murine tumor cell lines. Jpn J Exp Med., 59: 157~161, 1989.
7. Chiu JH, Ju CH., Wu LH, Lui WY, Wu CW, Shiao MS, Hong CY. Cordyceps sinensis increases the expression of major histocompatibility complex class II antigens on human hepatoma cell line HA22T/VGH cells. Am J Chin Med., 26: 159~170, 1998.
8. Nakamura K, Yamaguchi Y, Kagota S, Kwon YM, Shinozuka K, Kunitomo M. Inhibitory effect of Cordyceps sinensis on spontaneous liver metastasis of Lewis lung carcinoma and B16 melanoma cells in syngeneic mice. Jpn J Pharmacol., 79: 334~341, 1999.

9. Chen GZ, Chen G L, Sun T, Hsieh GC, and Henshall JM. Effects of Cordyceps sinensis on murine T lymphocyte subsets. Chin Med J (Engl), 104: 4-8, 1991.
10. Gao Q, Wu G, and He D. Effect of Cordyceps sinensis on the Th1/Th2 cytokines in patients with Condyloma Acuminatum. Zhong Yao Cai, 23: 402-404, 2000.
11. Gong HY, Wang KQ, and Tang SG. Effects of cordyceps sinensis on T lymphocyte subsets and hepatofibrosis in patients with chronic hepatitis B. Hunan Yi Ke Da Xue Xue Bao, 25: 248-250, 2000.
12. Guan YJ, Hu Z and Hou M. Effect of Cordyceps sinesis on T-lymphocyte subsets in chronic renal failure. Zhongguo Zhong Xi Yi Jie He Za Zhi, 12: 338-339, 323, 1992.
13. Wani MC, Taylor HL, Wall ME, et al. The isolation and structure of taxol, a novel antileukemic and antitumor agent from Taxus brevifolia. Journal of the American Chemical Society, 93: 2325-2327, 1971.
14. Rowinsky EK. and Donehower RC. Paclitaxel (taxol). N Engl J Med, 332: 1004-1014, 1995.
15. Manfredi JJ. and Horwitz SB. Taxol: an antimitotic agent with a new mechanism of action. Pharmacol Ther, 25: 83-125, 1984.
16. Holmes FA, Walters RS, Theriault RL, et al. Phase II trial of taxol, an active drug in the treatment of metastatic breast cancer. J Natl Cancer Inst, 83: 1797-1805, 1991.
17. Murphy WK, Fossella FV, et al. Phase II study of taxol in patients with untreated advanced non-small-cell lung cancer. J Natl Cancer Inst, 85: 384-388, 1993.
18. Ettinger DS, Finkelstein DM, Sarma RP, and Johnson DH. Phase II study of paclitaxel in patients with extensive-disease small-cell lung cancer: an Eastern Cooperative Oncology Group study. J Clin Oncol, 13: 1430-1435, 1995.
19. Forastiere AA, Neuberg D, Taylor SG, DeConti R, and Adams G. Phase II evaluation of Taxol in advanced head and neck cancer: an Eastern Cooperative Oncology group trial. J Natl Cancer Inst Monogr: 181-184, 1993.
20. Neri B, Cipriani G, Fulignati C, et al. Weekly paclitaxel and epirubicin in the treatment of symptomatic hormone-refractory advanced prostate carcinoma: report of a phase II trial. Anticancer Drugs, 16: 63-66, 2005.
21. Rowinsky EK, Eisenhauer EA, Chaudhry V, Arbuck SG, and Donehower RC. Clinical toxicities encountered with paclitaxel (Taxol). Semin Oncol, 20: 1-15, 1993.
22. Benjamin S, Merav W, Gennadi K, et al. Ammonium Trichloro (dioxoethylene-o,o') tellurate (AS101) Sensitizes Tumors to Chemotherapy by Inhibiting the Tumor Interleukin 10 Autocrine Loop. Cancer Res., 64: 1843-1852, 2003.
23. Tishler, R. B., Geard, C. R., Hall, E. J., and Schiff, P. B. Taxol sensitizes human astrocytoma cells to radiation. Cancer Res, 52: 3495-3497, 1992.
24. Chang YF, Li LL, Wu CW, et al. Paclitaxel-induced apoptosis in human gastric carcinoma cell lines. Cancer, 77: 14-18, 1996.
25. Speicher LA, Barone L, and Tew KD. Combined antimicrotubule activity of estramustine and taxol in human prostatic carcinoma cell lines. Cancer Res, 52: 4433-4440, 1992.
26. Stearns ME. and Wang M. Taxol blocks processes essential for prostate tumor cell (PC-3 ML) invasion and metastases. Cancer Res, 52: 3776-3781, 1992.
27. Zhang W, Yang J, Chen J, Hou Y, and Han X. Immunomodulatory and antitumor effects of exopolysaccharide fraction (EPSF) from a cultivated Cordyceps sinensis fungus on tumor-bearing mice. Biotechnol Appl Biochem, 2004.
28. Janeway CA. Travers P, Walport M, Shlomchik M. Immunobiology. New York and London: Garland Publishing, 2001.
29. Kalyuzhny AE. Chemistry and biology of the ELISPOT assay. Methods Mol Biol, 302: 15-32, 2005.
30. Shin KH, Lim SS, Lee S, Lee YS, Jung SH, and Cho SY. Anti-tumour and immuno-stimulating activities of the fruiting bodies of Paecilomyces japonica, a new type of Cordyceps spp. Phytother Res, 17: 830-833, 2003.
31. Koh JH, Yu KW, Suh HJ, Choi YM, and Ahn TS. Activation of macrophages and the intestinal immune system by an orally administered decoction from cultured mycelia of Cordyceps sinensis. Biosci Biotechnol Biochem, 66: 407-411, 2002.

全文公開日期本全文未授權公開 (校內網路)
全文公開日期本全文未授權公開 (校外網路)
全文公開日期本全文未授權公開 (國家圖書館：臺灣博碩士論文系統)

簡易檢索 / 詳目顯示

相關論文