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研究生:陳洳君
研究生(外文):Chen. Ju-Chun
論文名稱:靈芝、樟芝、椎茸、桑黃萃取物在調節免疫反應及抗B型肝炎病毒能力之研究第一部份:BLAB/c小鼠口服後免疫功能調節之研究第二部份:invitro抗B型肝炎病毒能力之研究
論文名稱(外文):Study of Ganoderma lucidum, Antrodia camphorata, Shiitake mushroom, Phellinus linteus extracts in immune responses modulation and anti-HBV capability
指導教授:黃惠宇黃惠宇引用關係
指導教授(外文):Hui-Yu Huang
學位類別:碩士
校院名稱:實踐大學
系所名稱:食品營養研究所
學門:醫藥衛生學門
學類:營養學類
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:116
中文關鍵詞:免疫調節靈芝樟芝椎茸桑黃抗B型肝炎病毒
外文關鍵詞:immunityGanoderma lucidumAntrodia camphorataShiitake mushroomPhellinus linteusanti-HBV
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「醫食同源」,這句名言是菇類作為食品的最佳註解。 在先前的文獻指出,菇類的萃取物擁有許多藥用的特性,不論是在體內或是在體外的測試中皆可以加強各種免疫反應。 因此本篇研究的目的,是想要評估長期口服之方式餵食靈芝、樟芝、椎茸和桑黃這種四種菇類是否會影響年輕BLAB/c小鼠的免疫反應,此外,實驗當中也會同時評估四種菇類之熱水萃取物抑制HepES-2肝癌細胞分泌B型肝炎病毒之能力與菇類熱水萃取物之抗氧化活性。
本篇研究中動物活體外測試的部分,是以三至四週齡大的BLAB/c雄性小白鼠為實驗動物,使用灌食的方式分別給予靈芝、樟芝、椎茸及桑黃四種菇類,劑量由低至高分別為20 mg/kg小鼠體重/day、100 mg/kg小鼠體重/day及250 mg/kg小鼠體重/day,個別餵食8週後,將老鼠犧牲並取出脾臟細胞作測試。 以MTT assay分析脾臟淋巴細胞受刺激後之細胞增生率; 再以流式細胞儀評估脾臟淋巴細胞受刺激後T、B淋巴細胞及NK自然殺手細胞增生的改變量; 最後利用酵素免疫測定法 (ELISA)偵測脾臟淋巴細胞在體外受刺激產生細胞激素IL-2、IL-4、IFN-γ的變化量。
結果顯示,長期給予靈芝餵食之小鼠,雖然脾臟淋巴細胞受到外來抗原刺激後其增生率並無顯著差異,但由流式細胞儀結果得知T淋巴細胞、NK自然殺手細胞之細胞數與由ELISA測得之細胞激素IL-2、IL-4的分泌量上,與控制組比較皆有顯著上升。 在長期給予樟芝餵食之小鼠中,脾臟淋巴細胞受到外來抗原刺激後其增生率並無顯著差異,但由流式細胞儀結果得知T淋巴細胞、NK自然殺手細胞之細胞數與由ELISA測得之細胞激素IFN-γ的分泌量上,與控制組比較皆有顯著上升的差異存在。 而在長期給予椎茸餵食之小鼠中,脾臟淋巴細胞受到外來抗原刺激後其增生率有顯著的上升,並且由流式細胞儀結果得知T、B淋巴細胞、NK自然殺手細胞之細胞數也明顯的增加,另外在由ELISA測得之細胞激素IL-2、IL-4、IFN-γ的分泌量上,與控制組比較有顯著上升的現象。 另外,在桑黃的實驗中,長期給予小鼠桑黃之餵食,會使得其脾臟淋巴細胞受到外來抗原刺激後增生率有顯著上升,並且由流式細胞儀結果得知T、B淋巴細胞增生之細胞數和ELISA測得之細胞激素IL-2、IL-4、IFN-γ的分泌量上,與控制組比較亦有顯著上升。
因此由本實驗結果推論,長期口服靈芝及樟芝在受到外來抗原刺激後,可以使T淋巴細胞和NK自然殺手細胞活性增加,而長期口服椎茸及桑黃在受到外來抗原刺激後則可以使T、B淋巴細胞和NK自然殺手細胞活性增加。 因此,宿主長期口頭服用靈芝、樟芝、椎茸和桑黃這四種菇類皆可以增強健康宿主的免疫能力。
本篇研究中細胞實驗的部分,則以HepG2細胞轉殖HBV基因而成的HepES-2細胞株(具有分泌HBe抗原及HBs抗原的能力)為測試目標,使用四種菇類冷凍乾燥後的熱水萃取物處理細胞,以MTT assay測試菇類熱水萃取物對HepES-2細胞的毒性劑量,以選取最適合之測試濃度 (對細胞毒性小於15%) 。 以5×104個的HepES-2細胞量,利用酵素免疫測定法(ELISA)偵測HepES-2細胞分泌e抗原及s抗原的分泌量,最後以清除DPPH自由基能力的測試來評估四種菇類熱水萃取物之抗氧化能力。
結果顯示,MTT assay測試中靈芝、樟芝及椎茸只需要125μg/ml的劑量處理後即發現對HepES-2細胞有明顯的毒性,但桑黃則無此現象。 而在B型肝炎病毒抗病毒方面,這四種菇類與控制組比較皆擁有明顯抑制HepES-2細胞分泌核心抗原e抗原之能力,但只有靈芝及椎茸擁有顯著抑制表面抗原s抗原的能力。 此外,這四種菇類的萃取物所擁有之清除DPPH自由基的能力是隨著劑量的增加而上升 (250 ~ 2000μg/ml)。 因此由本實驗結果推論,靈芝及椎茸的熱水萃取物擁有顯著抗B型肝炎病毒活性之能力,未來可以開發作為協助或使用於B型肝炎感染病人之治療用藥。
Mushrooms have been known as a medicine in traditional Chinese medicine. From previously studies, there are many result showed that the mushrooms extracts could enhanced immunity response both on in vivo and in vitro model. The aim of this study, we try to investigate whether long term oral administration Ganoderma lucidum, Antrodia camphorata, Shiitake mushroom, and Phellinus linteus can effect the immunity for young mice. Besides, we also would like to explore the inhibition of those mushrooms extract on hepatitis B virus (HBV) with HepES-2 cells, and do those extracts have antioxidant activity.
In animal model, the 3 to 4 weeks old male BLAB/c mice were divided into 4 groups oral administered 4 kinds of mushrooms G. lucidum, A. camphorata, S. mushroom and P. linteus at different dose age, 0 mg /Kg bw /day, 20 mg /Kg bw /day, 100 mg /Kg bw /day, and 250 mg /Kg bw /day, respectively for 8 weeks. MTT assay was used for detection of the lymphocyte proliferation rate. The flow cytometry analysis was used to estimate T, B, and NK cell number. The ELISA assay was used to determine cytokine (IL-2, IL-4, IFN- γ) secretion.
The result showed that long term oral administration G. lucidum in BLAB/c mice could increased T, NK cell numbers (by using flow cytometry), IL-2 and IL-4 secretion (by using ELISA) when compared with control group. The lymphocyte proliferation rate was no significant difference in MTT assay, but T, NK cell numbers (by using flow cytometry) and IFN-γsecretion (by using ELISA) had significant increase with long term oral administration of A. camphorata when compared with control group in BLAB/c mice . On the other hand, long term oral administration S. mushroom in BLAB/c mice, the lymphocyte proliferation rate had significant increase in MTT assay, T, B, NK cell numbers (by using flow cytometry), IL-2, IL-4 and IFN-γsecretion (by using ELISA) also had significant increase when compared with control group. The lymphocyte proliferation rate was significant increase in MTT assay, T, B cell numbers (by using flow cytometry), IL-2, IL-4 and IFN-γsecretion (by using ELISA) also had significant increase with long term oral administration of P. linteus when compared with control group in BLAB/c mice.
In this study, we conclude that long term oral administration G. lucidum and A. camphorata might enhanced T and NK cell activation, proliferation after antigen stimulation, and long term oral administration S. mushroom and P. linteus might enhanced T、B and NK cell activation, proliferation after antigen stimulation. Thus, long term oral administration mushrooms in mice could enhance the immunity of health host when they exposed in antigen.
HepES-2 cells, which derived from HepG2 cells, transfected with HBV genome, and HepES-2 cells has been examined with constant HBeAg and HBsAg expression. The hot water extract of those mushrooms were used to treat with HepES-2 cells, and MTT assay was used to detect the cytotoxcity of those mushroom extracts. HepES-2 cells were plated at a density of 5×104 cells with suitable concentration of mushrooms extract, which caused less than 15 % HepES-2 cells death. The secretion of HBeAg and HBsAg from HepES-2 cells were determined via ELISA assay. DPPH (1, 1-diphenyl-2-picrylhydrazyl) assay was used for evaluation of antioxidant activity of those mushroom extracts.
The result showed that G. lucidum, A. camphorata, and S. mushroom extracts had significant cytotoxcity for HepES-2 cells start from the concentration of 125μg/ml treatment. Compared with those in sense control, all mushrooms have significant decrease secretion of HBeAg from HepES-2 cells, but only G. lucidum and S. mushroom have significant decrease secretion of HBsAg from HepES-2 cells. Besides, those extract of mushrooms scavenged the stable free radical DPPH resulting in a increase of does-dependently (250 ~ 2000μg/ml). Thus G. lucidum and S. mushroom extract possesses a significant anti-HBV activity and perhaps can develop into adjuvant or therapeutic drug of HBV infection patients in the future.
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論文大綱
中文摘要 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 01
英文摘要 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 03
壹. 緒言 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 06
一. 文獻回顧 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 06
(一) 菇類之介紹 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 07
1. 靈芝 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 07
2. 樟芝 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 11
3. 椎茸 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 14
4. 桑黃 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 16
(二) 菇類功能之研究 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 19
1. 抗腫瘤、癌症之功能 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 19
2. 免疫力之調節 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 22
3. 抗細菌、微生物及病毒之功能 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 26
4. 保肝之作用 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 27
5. 抗氧化、抗發炎之功能 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 29
6. 其他能力:如抗突變能力、降血糖、降血脂 ‥‥‥‥‥‥‥‥‥‥ 31
(三) 中草藥對B型肝炎的治療方式 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 33
(四) 實驗設計之原理-免疫機轉 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥34
二. 本實驗之研究動機 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 38
(一) 菇類對BLAB/c小鼠免疫功能調節之影響 ‥‥‥‥‥‥‥‥‥‥‥ 38
(二) 菇類對抗B型肝炎病毒之能力 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 38
(三) 研究架構 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 39

貳. 材料與方法 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 40
一. 實驗材料 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 40
(一) 實驗動物 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 40
(二) 細胞株 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 40
(三) 樣品 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 40
1. 動物實驗部分(in vivo model) ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 41
2. 細胞實驗部分(in vitro model) ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 41
(四) 藥品及試劑 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 42
1. 動物實驗部分 (in vivo model) ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 42
2. 細胞實驗部分 (in vitro model) ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 43
二. 實驗方法 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 44
(一) 動物實驗部分 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 44
1. 分組 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 44
2. 動物犧牲及採檢體 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 44
3. 脾臟淋巴細胞增生測試MTT assay ‥‥‥‥‥‥‥‥‥‥‥‥‥ 44
4. 脾臟細胞中淋巴細胞增生數目之分析 ‥‥‥‥‥‥‥‥‥‥‥‥ 45
5. 脾臟淋巴細胞細胞激素分泌量之分析 ‥‥‥‥‥‥‥‥‥‥‥‥ 45
(二) 細胞實驗部分 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 46
1. 樣品配製 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 46
2. 細胞繼代培養 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 46
3. 細胞毒性測試MTT assay ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 47
4. 抗B型肝炎病毒之測試Anti-virus test(HBe Ag、HBs Ag)‥‥ 47
5. 清除DPPH自由基能力測試 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 48
三. 方法原理介紹 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 48
(一) Flow cytometry (流式細胞儀) ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 48
(二) MTT assay ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 49
(三) ELISA assay (酵素免疫測定法) ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 50
(四) DPPH antioxidant assay (清除DPPH自由基能力測試) ‥‥‥‥‥‥ 50
四. 統計分析 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 51

參. 結果 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 52
一. 餵食靈芝、樟芝、椎茸、桑黃對BLAB/c小鼠體重之影響 ‥‥‥‥‥‥‥ 52
二. 菇類對脾臟細胞增生能力之影響 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 52
(一) 餵食靈芝對脾臟淋巴細胞增生能力之影響 ‥‥‥‥‥‥‥‥‥‥‥ 52
(二) 餵食樟芝對脾臟淋巴細胞增生能力之影響 ‥‥‥‥‥‥‥‥‥‥‥ 53
(三) 餵食椎茸對脾臟淋巴細胞增生能力之影響 ‥‥‥‥‥‥‥‥‥‥‥ 53
(四) 餵食桑黃對脾臟淋巴細胞增生能力之影響 ‥‥‥‥‥‥‥‥‥‥‥ 54
三. 餵食菇類對脾臟細胞中淋巴細胞群增生數量之影響 ‥‥‥‥‥‥‥‥‥ 56
(一) 餵食靈芝對脾臟細胞中淋巴細胞群增生數量之影響 ‥‥‥‥‥‥‥ 56
(二) 餵食樟芝對脾臟細胞中淋巴細胞群增生數量之影響 ‥‥‥‥‥‥‥ 56
(三) 餵食椎茸對脾臟細胞中淋巴細胞群增生數量之影響 ‥‥‥‥‥‥‥ 57
(四) 餵食桑黃對脾臟細胞中淋巴細胞群增生數量之影響 ‥‥‥‥‥‥‥ 57
四. 餵食菇類對脾臟淋巴細胞細胞激素分泌量之影響 ‥‥‥‥‥‥‥‥‥‥ 58
(一) 餵食靈芝對脾臟淋巴細胞細胞激素分泌量之影響 ‥‥‥‥‥‥‥‥ 58
(二) 餵食樟芝對脾臟淋巴細胞細胞激素分泌量之影響 ‥‥‥‥‥‥‥‥ 59
(三) 餵食椎茸對脾臟淋巴細胞細胞激素分泌量之影響 ‥‥‥‥‥‥‥‥ 60
(四) 餵食桑黃對脾臟淋巴細胞細胞激素分泌量之影響 ‥‥‥‥‥‥‥‥ 61
五. 菇類對肝癌細胞HepES-2之毒性分析 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 62
(一) 靈芝對肝癌細胞HepES-2之毒性分析 ‥‥‥‥‥‥‥‥‥‥‥‥‥ 62
(二) 樟芝對肝癌細胞HepES-2之毒性分析 ‥‥‥‥‥‥‥‥‥‥‥‥‥ 62
(三) 椎茸對肝癌細胞HepES-2之毒性分析 ‥‥‥‥‥‥‥‥‥‥‥‥‥ 62
(四) 桑黃對肝癌細胞HepES-2之毒性分析 ‥‥‥‥‥‥‥‥‥‥‥‥‥ 63
六. 菇類抗B型肝炎病毒能力之評估 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 64
(一) 菇類抗B型肝炎病毒e抗原能力之評估 ‥‥‥‥‥‥‥‥‥‥‥‥ 64
(二) 菇類抗B型肝炎病毒s抗原能力之評估 ‥‥‥‥‥‥‥‥‥‥‥‥ 65
七. 菇類之抗氧化能力分析 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 66

結果圖表 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 68

肆. 討論 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 83

伍. 結論 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 104

陸. 參考文獻 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 109

表 目 錄
表1-1 靈芝之一般化學成分 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 09
表2-1 chow diet 5010 (PMI) 其化學組成 ‥‥‥‥‥‥‥‥‥‥‥‥‥ 41
表3-1 利用MTT法偵測餵食靈芝對脾臟淋巴細胞增生能力之影響 ‥‥‥‥ 69
表3-2 利用MTT法偵測餵食樟芝對脾臟淋巴細胞增生能力之影響 ‥‥‥‥ 70
表3-3 利用MTT法偵測餵食椎茸對脾臟淋巴細胞增生能力之影響 ‥‥‥‥ 71
表3-4 利用MTT法偵測餵食桑黃對脾臟淋巴細胞增生能力之影響 ‥‥‥‥ 72
表3-5 利用流式細胞儀偵測餵食靈芝、樟芝對脾臟細胞中淋巴細胞群增生數量之影響 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 73
表3-6 利用流式細胞儀偵測餵食椎茸、桑黃對脾臟細胞中淋巴細胞群增生數量之影響 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 74
表3-7 利用ELISA reader偵測餵食靈芝對脾臟淋巴細胞細胞激素分泌量之影響 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 75
表3-8 利用ELISA reader偵測餵食樟芝對脾臟淋巴細胞細胞激素分泌量之影響 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 76
表3-9 利用ELISA reader偵測餵食椎茸對脾臟淋巴細胞細胞激素分泌量之影響 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 77
表3-10 利用ELISA reader偵測餵食桑黃對脾臟淋巴細胞細胞激素分泌量之影響 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 78
表3-11 菇類熱水萃取物對肝癌細胞HepES-2之毒性分析 ‥‥‥‥‥‥‥‥ 79
表3-12 菇類熱水萃取物抑制肝癌細胞HepES-2分泌e抗原之能力分析 ‥‥ 80
表3-13 菇類熱水萃取物抑制肝癌細胞HepES-2分泌s抗原之能力分析 ‥‥ 81
表5-1 口服不同菇類對BLAB/c小鼠體重及脾臟淋巴細胞影響之綜合整理 104
表5-2 菇類熱水萃取物對肝癌細胞HepES-2之影響及其抗氧化能力分析之綜合整理 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 106

圖 目 錄
圖1-1 靈芝之型態外觀 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 07
圖1-2 靈芝之高分子多醣體分子結構圖 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 10
圖1-3 靈芝酸之分子結構圖 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 10
圖1-4 樟芝之型態外觀 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 11
圖1-5 椎茸之型態外觀 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 14
圖1-6 桑黃之型態外觀 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 16
圖1-7 實驗設計之原理-免疫機轉 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 37
圖2-1 流式細胞儀之操作原理 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 49
圖2-2 酵素免疫測定法之操作原理 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 50
圖3-1 餵食菇類對BLAB/c小鼠體重之影響 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 68
圖3-2 菇類熱水萃取物清除DPPH自由基能力之分析 ‥‥‥‥‥‥‥‥‥‥ 82
圖5-1 實驗設計之原理: 菇類作用之免疫機轉 ‥‥‥‥‥‥‥‥‥‥‥‥ 108




縮寫對照表

縮寫字 原文或原意
T cell T lymphocyte
B cell B lymphocyte
NK cell Nature killer cell
Con A Concanavalin A
LPS Lipopolysaccharide
Th1 Type 1 T helper cell
Th2 Type 2 T helper cell
IL - 2 Interleukin - 2
IL - 4 Interleukin - 4
IFN -γ Interferon -γ
MTT 3-(4,5-dimethyl thiazol-2-yl) -2,5-diphenyl-tetrazolium bromide
FBS Fetal bovine serum
DMEM Dulbecco’s modified Eagle’s medium
BSA Bovine serum albumin
PBS Phosphate-buffered saline
DPPH 1,1-diphenyl-2-picryl-hydrazyl
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1. 12、陳愛娥,「訴訟權能」與「訴訟利益」──從兩件行政法院裁判出發,觀察兩種訴訟要件的意義與功能,律師雜誌第二五四期十一月號,民國89年11月。
2. 11、陳清秀,新行政訴訟種類之評析與展望,月旦法學雜誌第四十七期,民國88年4月。
3. 4、陳敏,租稅法之連帶債務,政大法學評論第二十八期,民國72年12月。
4. 10、陳清秀,稅法上責任債務(第二次納稅義務),東吳法律學報第八卷第二期,民國84年3月。
5. 9、陳清秀,稅法上之連帶債務,財稅研究二十六卷第二期,民國83年3月。
6. 7、陳敏,租稅行政處分之通知方式,政大法學評論第四十七期,民國82年6月。
7. 5、陳敏,扣繳薪資所得税之法律關係,政大法學評論第五十一期,民國83年6月。
8. 2、吳庚,行政訴訟中各類之關係,法令月刊四十七卷第十一期,民國87年11月。
9. 13、黃俊杰,中科院之扣繳義務人處罰之爭議,月旦法學雜誌九十三期,民國93年2月。
10. 14、黃茂榮,稅捐債務之繳納義務人,經社法制論叢第二期,民國77年7月。
11. 5、水野武夫,鄭俊仁譯,第二次納稅義務(上),植根雜誌第十卷第七期,民國83年7月。
12. 吳庚,〈近年國內公法的發展趨勢〉,《法學講座》,第12期(2002年12月)
13. 李建良,〈行政程序法與正當法律程序 ─初論行政程序法的立法目的與規範內涵〉,《憲政時代》,第25卷第3期(2000年1月)
14. 李建良,〈基本權利的理念變遷與功能體系─從耶林內克「身份理論」談起(下)〉,《憲政時代》,第29卷第2期(2003年10月)
15. 李建良,〈論基本權利之程序功能與程序基本權─德國理論的借鑑與反思〉,《憲政時代》,第29卷第4期,頁527(2004年4月)
 
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