探討化學治療後,餵食冬蟲夏草對小鼠免疫機制的影響

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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：2 下載：0
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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

                                

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