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  1. NTU Theses and Dissertations Repository
  2. 生物資源暨農學院
  3. 園藝暨景觀學系
Please use this identifier to cite or link to this item: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25346
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???org.dspace.app.webui.jsptag.ItemTag.dcfield???ValueLanguage
dc.contributor.advisor許輔(Fuu Sheu)
dc.contributor.authorYing-Yi Chenen
dc.contributor.author陳盈宜zh_TW
dc.date.accessioned2021-06-08T06:09:54Z-
dc.date.copyright2007-07-27
dc.date.issued2007
dc.date.submitted2007-07-11
dc.identifier.citation參考文獻
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25346-
dc.description.abstract第一部份以山防風(Echinops grijsii)、山葡萄(Ampelopsis brevipedunculata Maxim)、白鶴靈芝 (Rhinacanthus nasutus L. Kurz)、荔枝草 (Salvia plebeia R. Brown) 及雞血藤 (Millettia reticulata) 之乾燥粉末,做成複方中草藥配方 A 與 B 並以BALB/c 小鼠為對象,探討其個別之免疫調節活性。非特異性免疫活性結果顯示餵食複方 A 高劑量 (5.4 mg/mouse) 與複方 B 高劑量 (4 mg/mouse) 可顯著提高小鼠脾細胞增生率;餵食複方 A 高、中 (1.8 mg/mouse)、低 (0.6 mg/mouse) 三劑量與複方 B 高及中 (1.3 mg/mouse) 劑量均能顯著提升脾細胞 IFN-γ 之分泌量,餵食複方 A 中劑量與複方 B 高劑量亦能提高脾細胞 IL-2 之分泌量;而餵食複方A 與複方 B 三劑量,相較於 PBS 組可提高小鼠脾細胞中 NK cells 之活性;餵食複方 A 中、低劑量與複方 B 高、中劑量之血液中單核球細胞的吞噬能力皆顯著高於餵食 PBS 組;餵食複方 A 高劑量會顯著增加 TH 細胞且中劑量會顯著增加B cell 之比例,而複方 B 三劑量則無顯著差異。餵食複方 A 高、中劑量與複方B 高、中、低 (0.5 mg/mouse) 三劑量均能顯著提升小鼠血清中 IFN-γ 及 IL-2 之分泌量,此外,餵食複方 B 高、低劑量之小鼠,相較於餵食對照組 (PBS) 能顯著提高血清中總 IgG 濃度。
OVA 特異性免疫活性結果顯示餵食複方 A 與複方 B 三劑量均能顯著提高小鼠 OVA 特異性之脾細胞增生率;而餵食複方 A 與複方 B 高及中劑量均能顯著提升 OVA 特異性 IFN-γ 之分泌量,餵食複方 B 高、中、三劑量則可顯著提升 OVA 特異性 IL-2 之分泌量。此外,餵食複方 A 與複方 B 高、中劑量相較於餵食對照組 (PBS) 能顯著提高血清中 OVA 特異性之 IgG 濃度,有助於體液性免疫反應而防禦對抗侵害性之特異抗原。上述結果說明餵食複方中草藥配方 A與配方 B 於高及中劑量均有助於提高小鼠的非特異性與 OVA 特異性免疫反應,具醫藥或健康食品之應用潛力。
第二部份主要探討樟芝 (Antrodia cinnamomea) 子實體的抗腫瘤活性及其機制。乾燥之樟芝子實體粉末以去離子水加熱煮沸配製懸浮水溶液,以低 (0.22 mg/mouse)、中 (0.66 mg/mouse) 及高劑量 (2.00 mg/mouse) 為試驗組,並以去離子水 (deionized water, DI) 作為對照組,以及本實驗室純化所得之金針菇免疫調節蛋白 FIP-fve (0.20 mg/mouse) 作為正控制組。罹癌小鼠存活實驗顯示,注射 ATCC BNL 1MEA.7R.1 肝癌細胞 (3x104 cells/mouse) 之 BALB/c 小鼠在餵食高、中劑量樟芝樣本後,相較餵食對照組 (DI),可顯著延長罹癌小鼠存活時間分別達 47.4 % 及 57.1 %。
在非特異性抗腫瘤活性方面,結果顯示餵食樟芝子實體水萃物高中低三劑量組之小鼠,均可顯著活化腹腔巨噬細胞提升細胞激素 TNF-α 分泌量及 NO 之產生量;且小鼠腹腔巨噬細胞毒殺小鼠肝癌細胞能力,與對照組相比皆具有顯著性差異。在腫瘤特異性抗腫瘤活性方面,結果顯示餵食樟芝子實體高中低三劑量均可顯著促進腫瘤特異性之小鼠脾細胞增生作用;並顯著提升小鼠脾細胞之腫瘤特異 IFN-γ 分泌量及顯著提高腫瘤特異性 IgG 分泌量;且小鼠脾臟細胞毒殺小鼠肝癌細胞能力,與對照組相比皆具有顯著性差異;而餵食高及中劑量組可顯著提升小鼠脾細胞之腫瘤特異性 TNF-α 及 IL-2 分泌量。以上結果顯示餵食樟芝子實體於中高劑量,均具有活化非特異性及腫瘤特異性之免疫調節活性,因而具有抑制小鼠肝腫瘤之作用。
zh_TW
dc.description.abstractIn part І, the objective was to evaluate the immunomodulatory effects of Chinese herbal formula A (CHFA) and Chinese herbal formula B (CHFB). The results of the evaluation of non-specific immunomodulatory activities showed that administrating of high doses of CHFA (5.4 mg/mouse) and CHFB (4.0 mg/mouse) significantly (p < 0.05) enhanced cell proliferation of mouse splenocytes. Administration of CHFA at high, medium (1.8 mg/mouse), and low (0.6 mg/mouse) doses and CHFB at high, and medium (1.3 mg/mouse) doses significantly (p < 0.05) increased IFN-γ secretion by mouse splenocytes. Administration of CHFA (medium dose) and CHFB (high dose) significantly (p < 0.05) increased IL-2 secretion by mouse splenocytes. All three doses of CHFA and CHFB significantly (p < 0.05) enhanced the NK cells activity of mouse splenocytes. Administration of CHFA and CHFB at high and medium doses significantly (p < 0.05) enhanced the phagocytosis activities of monocytes. High and medium doses of CHFA significantly (p < 0.05) increased the ratio of TH cells and B cells in the mouse splenocytes. Administration of CHFA (high and medium doses) and CHFB high, medium, and low (0.5 mg/mouse) doses significantly (p < 0.05) increased the secretion of IFN-γ and IL-2 in sera. Taking high and low doses of CHFB significantly (p < 0.05) increased total IgG secretion in sera.
The results of the evaluation on OVA-specific immunomodulatory activities showed that administration of CHFA and CHFB at high, medium, and low doses significantly (p < 0.05) enhanced the OVA-specific cell proliferation of mouse splenocytes. The formulas at high and medium doses significantly (p < 0.05) increased OVA-specific IFN-γ secretion by mouse splenocytes. The consumption of CHFB at all three doses significantly (p < 0.05) increased OVA-specific IL-2 secretion by mouse splenocytes. Moreover, high and medium doses of the two formulas significantly (p < 0.05) enhanced OVA-specific IgG secretion in sera. These results demonstrated that administration of CHFA and CHFB had immunomodulatory functions. Both these formulas might have potentials in related medicinal and health food applications.
In part Ⅱ, the objective was to investigate the anti-tumor effects and its related mechanisms of Antrodia cinnamomea fruiting bodies. Oral administration of high (2.00 mg/mouse) and medium (0.66 mg/mouse) doses of A. cinnamomea fruiting body samples significantly (p < 0.05) increased the life span of ATCC BNL 1MEA.7R.1 hepatoma-bearing mice (3×104 cells/mouse) by 47.4 % and 57.1 %, respectively. This result suggested that A. cinnamomea displayed activities to suppress hepatoma growth in vivo.
To understand the mechanism corresponding to this anti-tumor activity, effects of A. cinnamomea on activating the non-specific and tumor-specific immunity of the host were further investigated. The results of non-specific immunomodulatory activities showed that administration of A. cinnamomea fruiting body samples at high, medium, and low (0.22 mg/mouse) doses significantly (p < 0.05) stimulated mouse peritoneal macrophages to secreted TNF-α and produced NO. Additionally, the tumoricidal activity of peritoneal cells was also significantly enhanced (p < 0.05). The results of tumor-specific immunomodulatory activities showed that taking A. cinnamomea samples at three different doses significantly (p < 0.05) enhanced the tumor-specific cell proliferation of hepatoma-bearing mice splenocytes. A. cinnamomea samples at all three doses significantly (p < 0.05) increased tumor-specific IFN-γ secretion by hepatoma-bearing mice splenocytes and significantly (p < 0.05) enhanced tumor-specific IgG secretion in serum. High and medium doses of A. cinnamomea samples significantly (p < 0.05) increased tumor-specific TNF-α and IL-2 secretion by hepatoma-bearing mice splenocytes. The tumoricidal activity of splenocytes cells obtained from hepatoma-bearing mice fed the A. cinnamomea samples was also significantly enhanced (p < 0.05).
These results showed that A. cinnamomea fruiting body samples carried out anti-tumor activity by activating both the non-specific and tumor-specific immunity of their host.
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dc.description.tableofcontents目錄
口試委員會審定書 i
誌謝 ii
中文摘要 iii
英文摘要 v
第一章 研究背景
第一節 前言 1
第二節 本研究所選用中草藥之相關研究 2
一、山防風 2
(一)、山防風的簡介 2
(二)、山防風的生理機能 3
二、山葡萄 3
(一)、山葡萄的簡介 3
(二)、山葡萄的生理機能 4
三、白鶴靈芝 5
(一)、白鶴靈芝的簡介 5
(二)、白鶴靈芝的生理機能 5
四、荔枝草 6
(一)、荔枝草的簡介 6
(二)、荔枝草的生理機能 7
五、雞血藤 7
(一)、雞血藤的簡介 7
(二)、雞血藤的生理機能 8
六、樟芝 8
(一)、樟芝的分類與形態 8
(二)、樟芝的活性成分 9
(三)、樟芝的生理機能 10
第三節 免疫調節作用 12
一、先天性免疫反應 13
二、後天性免疫反應 14
三、自然殺手細胞 16
四、細胞激素 17
五、免疫球蛋白 17
六、免疫調節與腫瘤抑制 19
第四節 複方的理念與應用 20
第五節 研究動機與架構 21
第二章 材料與方法
壹、複方中草藥於BALB/c小鼠體內免疫調節功能之研究 23
第一節 中草藥之萃取製備 23
一、材料 23
二、萃取製備 23
第二節 體內 (in vivo) 非特異性免疫調節活性分析 23
一、材料 24
二、餵食複方中草藥材料之試驗設計 26
三、對小鼠脾臟細胞之影響 26
(一)、小鼠脾臟細胞的分離與製備 26
(二)、細胞代謝活性分析 (MTT assay) 26
(三)、淋巴細胞增生試驗 (BrdU assay) 27
(四)、自然殺手細胞活性試驗 28
(五)、脾臟細胞表面標記分析 30
(六)、細胞激素IFN-γ之測定 31
(七)、細胞激素IL-2之測定 32
四、血液中單核球細胞之吞噬試驗 33
五、對小鼠血清中總IgG及細胞激素含量之影響 35
(一)、小鼠血清之取得 35
(二)、總IgG之測定 35
(三)、細胞激素IFN-γ之測定 36
(四)、細胞激素IL-2之測定 37
第三節 體內 (in vivo) 特異性免疫調節活性分析 38
一、材料 38
二、餵食複方中草藥材料之試驗設計 40
三、對小鼠脾臟細胞之影響 40
(一)、小鼠脾臟細胞的分離與製備 40
(二)、OVA特異性細胞代謝活性試驗 (MTT assay) 41
(三)、OVA特異性淋巴細胞增生試驗 (BrdU assay) 41
(四)、OVA特異性細胞激素IFN-γ之測定 42
(五)、OVA特異性細胞激素IL-2之測定 43
四、對小鼠血清中OVA特異性IgG之影響 44
(一)、小鼠血清之取得 44
(二)、OVA特異性之IgG測定 45
第四節 統計分析 46
貳、樟芝子實體抑制小鼠肝腫瘤之研究 47
第一節 罹癌小鼠餵食樟芝子實體之存活試驗 47
一、材料 47
二、餵食樟芝子實體之存活試驗設計 47
第二節 體內 (in vivo) 非特異性抗腫瘤活性分析 48
一、材料 48
二、餵食樟芝子實體之試驗設計 50
三、對小鼠腹腔巨噬細胞之影響 50
(一)、腹腔巨噬細胞之分離與製備 50
(二)、細胞激素TNF-α分泌量之測定 51
(三)、一氧化氮 (nitric oxide) 產量之測定 52
(四)、小鼠腹腔巨噬細胞毒殺BNL 1MEA.7R.1小鼠肝癌細胞試驗 53
第三節 體內 (in vivo) 特異性抗腫瘤活性分析 54
一、材料 54
二、餵食樟芝子實體之試驗設計 56
三、對小鼠脾臟細胞之影響 57
(一)、小鼠脾臟細胞之分離與製備 57
(二)、腫瘤特異性淋巴細胞增生試驗 (BrdU assay) 57
(三)、腫瘤特異性細胞激素TNF-α之測定 59
(四)、腫瘤特異性細胞激素IFN-γ之測定 60
(五)、腫瘤特異性細胞激素IL-2之測定 61
(六)、腫瘤特異性脾臟細胞毒殺BNL 1MEA.7R.1小鼠肝癌細胞試驗 63
四、對小鼠血清中腫瘤特異性IgG之影響 64
(一)、小鼠血清之取得 64
(二)、腫瘤特異性之IgG測定 64
第四節 統計分析 65
第三章 實驗結果
壹、複方中草藥於BALB/c小鼠體內免疫調節功能之研究 66
第一節 體內 (in vivo) 非特異性免疫調節活性分析 66
ㄧ、飼養期間體重及犧牲後各臟器重量變化 66
二、對小鼠脾臟細胞之影響 66
(一)、細胞代謝活性試驗 (MTT assay) 66
(二)、淋巴細胞增生試驗 (BrdU assay) 67
(三)、自然殺手細胞活性試驗 67
(四)、脾臟細胞表面標記分析 68
(五)、細胞激素IFN-γ之測定 68
(六)、細胞激素IL-2之測定 69
三、血液中單核球細胞之吞噬試驗 69
四、對小鼠血清中總IgG及細胞激素含量之影響 70
(一)、總IgG產生量之測定 70
(二)、細胞激素IFN-γ之測定 70
(三)、細胞激素IL-2之測定 71
第二節 體內 (in vivo) 特異性免疫調節活性分析 71
ㄧ、飼養期間體重及犧牲後各臟器重量變化 71
二、對小鼠脾臟細胞之影響 72
(一)、OVA特異性細胞代謝活性試驗 (MTT assay) 72
(二)、OVA特異性淋巴細胞增生試驗 (BrdU assay) 72
(三)、OVA特異性細胞激素IFN-γ之測定 73
(四)、OVA特異性細胞激素IL-2之測定 74
三、對小鼠血清中OVA特異性IgG含量之影響 75
貳、樟芝子實體抑制小鼠肝腫瘤之研究 76
第一節 餵食樟芝子實體對罹癌小鼠存活之影響 76
第二節 餵食樟芝子實體可活化小鼠體內非特異性免疫反應 76
ㄧ、對小鼠腹腔巨噬細胞之影響 76
(一)、細胞激素TNF-α分泌量之測定 77
(二)、一氧化氮 (nitric oxide) 產量之測定 77
(三)、小鼠腹腔巨噬細胞毒殺BNL 1MEA.7R.1小鼠肝癌細胞試驗 78
第三節 餵食樟芝子實體可活化罹癌小鼠體內腫瘤特異性免疫反應 79
一、對小鼠脾臟細胞之影響 79
(一)、腫瘤特異性淋巴細胞增生試驗 (BrdU assay) 79
(二)、腫瘤特異性細胞激素TNF-α之測定 80
(三)、腫瘤特異性細胞激素IFN-γ之測定 80
(四)、腫瘤特異性細胞激素IL-2之測定 81
(五)、腫瘤特異性脾臟細胞毒殺BNL 1MEA.7R.1小鼠肝癌細胞試驗 82
二、對小鼠血清中腫瘤特異性IgG含量之影響 82
第四章 討論 84
第五章 結論 89
參考文獻 90
圖與表 101
圖目錄
圖一、非特異性免疫調節餵食流程圖 101
圖二、餵食中草藥複方A與B對小鼠脾臟細胞代謝 (MTT assay) 之影響 102
圖三、餵食中草藥複方A與B對小鼠脾臟細胞增生 (BrdU assay) 之影響 103
圖四、餵食中草藥複方A與B對小鼠脾臟中自然殺手細胞活性之影響 104
圖五、餵食中草藥複方A與B對小鼠血液中單核球細胞吞噬作用之影響 105
圖六、餵食中草藥複方A與B對小鼠脾臟細胞產生IFN-γ濃度之影響 106
圖七、餵食中草藥複方A與B對小鼠脾臟細胞產生IL-2濃度之影響 107
圖八、餵食中草藥複方A與B對小鼠血清中細胞激素IFN-γ濃度之測定 108
圖九、餵食中草藥複方A與B對小鼠血清中細胞激素IL-2濃度之測定 109
圖十、餵食中草藥複方A與B對小鼠血清中總IgG含量之影響 110
圖十一、特異性免疫調節餵食流程圖 111
圖十二、餵食中草藥複方A與B對經過OVA免疫之小鼠脾臟細胞產生OVA特異性細胞代謝活性 (MTT assay) 之影響 112
圖十三、餵食中草藥複方A與B對經過OVA免疫之小鼠脾臟細胞產生OVA特異性細胞代謝活性 (MTT assay) 百分比 113
圖十四、餵食中草藥複方A與B對經過OVA免疫之小鼠脾臟細胞產生OVA特異性細胞增生活性 (BrdU assay) 之影響 114
圖十五、餵食中草藥複方A與B對經過OVA免疫之小鼠脾臟細胞產生OVA特異性細胞增生活性 (BrdU assay) 百分比 115
圖十六、餵食中草藥複方A與B對經過OVA免疫之小鼠脾臟細胞產生OVA特異性IFN-γ之影響 116
圖十七、餵食中草藥複方A與B對經過OVA免疫之小鼠脾臟細胞產生OVA特異性IFN-γ百分比 117
圖十八、餵食中草藥複方A與B對經過OVA免疫之小鼠脾臟細胞產生OVA特異性IL-2之影響 118
圖十九、餵食中草藥複方A與B對經過OVA免疫之小鼠脾臟細胞產生OVA特異性IL-2百分比 119
圖二十、餵食中草藥複方A與B對小鼠血清中OVA特異性IgG含量之影響 120
圖二十一、小鼠餵食樟芝子實體流程圖 121
圖二十二、口服樟芝子實體對罹癌小鼠存活時間之影響 122
圖二十三、口服樟芝子實體對小鼠腹腔巨噬細胞分泌TNF-α之影響 123
圖二十四、口服樟芝子實體對小鼠腹腔巨噬細胞分泌NO之影響 124
圖二十五、口服樟芝子實體於小鼠腹腔巨噬細胞對BNL 1MEA.7R.1小鼠肝癌細胞之細胞毒殺活性影響 125
圖二十六、口服樟芝子實體對罹癌小鼠脾臟細胞產生腫瘤特異性細胞增生活性之影響 126
圖二十七、口服樟芝子實體對罹癌小鼠脾臟細胞產生腫瘤特異性細胞增生活性百分比 127
圖二十八、口服樟芝子實體對罹癌小鼠脾臟細胞產生腫瘤特異性TNF-α之影響 128
圖二十九、口服樟芝子實體對罹癌小鼠脾臟細胞產生腫瘤特異性IFN-γ之影響 129
圖三十、口服樟芝子實體對罹癌小鼠脾臟細胞產生腫瘤特異性IL-2之影響 130
圖三十一、口服樟芝子實體對罹癌小鼠產生腫瘤特異性IgG之影響 131
圖三十二、口服樟芝子實體於罹癌小鼠脾臟細胞對BNL 1MEA.7R.1小鼠肝癌細胞之腫瘤特異性細胞毒殺活性之影響 132
圖三十三、口服樟芝子實體於罹癌小鼠脾臟細胞中腫瘤特異性細胞激素與腫瘤特異性毒殺活性之相關性 133
表目錄
表一、餵食中草藥複方A之小鼠體重變化 134
表二、餵食中草藥複方A之小鼠各器官相對重量 135
表三、餵食中草藥複方B之小鼠體重變化 136
表四、餵食中草藥複方B之小鼠各器官相對重量 137
表五、餵食中草藥複方A對經過OVA免疫之小鼠體重變化 138
表六、餵食中草藥複方A對經過OVA免疫之小鼠各器官相對重量 139
表七、餵食中草藥複方B對經過OVA免疫之小鼠體重變化 140
表八、餵食中草藥複方B對經過OVA免疫之小鼠各器官相對重量 141
表九、餵食中草藥複方A小鼠脾臟細胞中淋巴細胞之比例 142
表十、餵食中草藥複方B小鼠脾臟細胞中淋巴細胞之比例 143
表十一、餵食中草藥複方A與B小鼠免疫反應之比較 144
表十二、餵食中草藥複方A與B對經過OVA免疫小鼠免疫反應之比較 145
表十三、口服樟芝子實體對罹癌小鼠存活率之影響 146
表十四、口服樟芝子實體抑制小鼠肝腫瘤之總結果 147
dc.language.isozh-TW
dc.subject免疫調節zh_TW
dc.subject複方中草藥zh_TW
dc.subject脾臟細胞zh_TW
dc.subject樟芝子實體zh_TW
dc.subject抗腫瘤zh_TW
dc.subject巨噬細胞zh_TW
dc.subjectsplenocyteen
dc.subjectmacrophageen
dc.subjectanti-tumor activityen
dc.subjectAntrodia cinnamomea fruiting bodyen
dc.subjectChinese herbal formulaen
dc.subjectimmunomodulatoryen
dc.title複方中草藥調節免疫及樟芝子實體抑制腫瘤之研究zh_TW
dc.titleStudies on the Immunomodulatory Properties of Chinese Herbal Formulas and the Anti-Tumor Effects of Antrodia cinnamomea Fruiting Bodiesen
dc.typeThesis
dc.date.schoolyear95-2
dc.description.degree碩士
dc.contributor.oralexamcommittee嚴國欽,林金源,陳中,周志輝
dc.subject.keyword複方中草藥,免疫調節,脾臟細胞,樟芝子實體,抗腫瘤,巨噬細胞,zh_TW
dc.subject.keywordChinese herbal formula,immunomodulatory,splenocyte,Antrodia cinnamomea fruiting body,anti-tumor activity,macrophage,en
dc.relation.page147
dc.rights.note未授權
dc.date.accepted2007-07-12
dc.contributor.author-college生物資源暨農學院zh_TW
dc.contributor.author-dept園藝學研究所zh_TW
Appears in Collections:園藝暨景觀學系

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