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研究生:謝如婷
研究生(外文):Ru-Ting Hsieh
論文名稱:微量元素對牛樟芝發酵物生理活性與抗氧化活性影響之探討
論文名稱(外文):Effects of Microelements on TheBioactivities and Antioxidant Activity of Antrodia camphorata Fermentation Product
指導教授:李順來李順來引用關係許孟博
指導教授(外文):Shun-Lai LiMeng-Bor Hsu
學位類別:碩士
校院名稱:南台科技大學
系所名稱:生物科技系
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:111
中文關鍵詞:牛樟芝抗氧化
外文關鍵詞:Antrodia camphorataAntioxidant Activity
相關次數:
  • 被引用被引用:6
  • 點閱點閱:1995
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:2
牛樟芝是目前最熱門的藥用菇菌,生理活性成份包含多醣體、三萜類等,其
藥理活性為抗腫瘤、抗氧化、保肝等相關研究相當多。文獻指出真菌於液態發酵
中添加某些微量元素會促使菌絲體胞內和胞外粗多醣含量增加及提昇菌體生長及
抗腫瘤活性,但對牛樟芝添加微量元素的影響則尚未有相關研究。因此,本研究
主要探討微量元素對牛樟芝發酵產物在多醣含量、抗氧化活性及體外試驗對腫瘤
細胞抑制能力的影響,並以HPLC 分析其發酵產物之極性分佈變化。
實驗結果顯示,液態發酵液部份,添加Se 可增加胞外多醣含量,添加微量元
素其總多酚含量均呈減少趨勢;添加Zn 及低濃度Mn 可提高清除DPPH 自由基能
力;添加1 ppm Mn、Fe 清除ABTS 自由基能力有明顯提昇;添加Ge、Mn 及低
濃度Zn 可提昇螯合亞鐵離子能力。若添加微量元素於中草藥牛樟芝液態發酵中,
添加Zn 及低濃度Fe 可增加胞外多醣含量;添加微量元素其總多酚含量呈上昇趨
勢,且DPPH 清除率及清除ABTS 能力亦提昇。腫瘤細胞測定部份,添加微量元
素於中草藥牛樟芝發酵液中,僅對抑制直腸癌細胞生長呈顯著增加。
液態菌絲體部份,添加微量元素隨著濃度的增加胞內多醣含量及總多酚含量
亦顯著增加。腫瘤細胞測定部份,添加微量元素對抑殺直腸癌細胞呈顯著增加,
添加Mn、Fe 對抑制肝癌細胞呈增加現象。
固態發酵產物方面,添加Mn 會增加其HPLC 分析低極性區域代謝物含量分
佈,但對中高極性化合物的影響不大;在含中草藥的固態發酵方面,發現1 ppm
Ge、Zn 及7.5 ppm Mn 可以增加發酵產物內的低極性代謝物含量分佈,但對中高
極性代謝物沒有影響。添加Fe 可增加發酵產物多醣含量的增加。添加Se、Ge 及
低濃度Mn、Zn 總多酚含量呈上昇現象;添加低濃度Mn、Zn 清除DPPH 自由基
能力亦增強;添加Se、Ge、Mn 可增加螯合亞鐵離子能力。中草藥牛樟芝固態發
酵中,添加微量元素其多醣含量無顯著性變化;添加Ge 及低濃度Se、Fe、Zn 可提昇總多酚含量及ABTS 清除能力;添加Ge、Mn 清除DPPH 自由基能力亦增強;
添加Mn、Fe 及低濃度Ge 螯合亞鐵離子能力呈增強現象。腫瘤細胞測定部份,添
加微量元素對抑制直腸癌細胞、肝癌細胞生長呈增加現象,另一方面,添加微量
元素於中草藥牛樟芝固態發酵產物亦是相同情形。總而言之,添加微量元素於牛
樟芝固態發酵產物中能改變菌絲體代謝物的含量及分佈,並可提昇其多醣含量、
抗氧化能力及抑制腫瘤細胞的生長。
Antrodia camphorata ( AC ) is the most popular medicinal mushroom in Taiwan. Its bioactive ingredients include olysaccharides,triterpenoids and so on. It has been reported to possess anti-tumor, antioxidant and hepatoprotective activities. Some research results indicated that addition of certain microelements into medium can enhance fungal mycelia growth, polysaccharide content and antitumor activity of the fermentation products. There is no related reports in AC was found so far. In this study microelements were used to investigate their effects on the polysaccharide content, antioxidant activity and the tumor cell inhibition capability of the fermentation products of AC. In addition, HPLC was used to analyze the influence of microelements on the polarity distrubution of the fermentation products.
The results of liquid-state fermented broth showed that Se elements can increase extracellular polysaccharides content, Zn and low concentration of Mn can enhance the DPPH scavenging effect while 1 ppm of Fe and Mn increase the ABTS scavenging activitives. Addition of Ge, Mn and low concentration of Zn can improve the chelating ability on ferrous ion of the fermented broth. The addition of microelements, however, will reduce the total phenol content of the broth. For the broth fermented with herbal medicines, the total phenol contents as well as the DPPH and ABTS scavenging capabilities were improved with microelement addition, while Zn and low concentration of Fe increased the level of extracellular polysaccharides content.
In the aspect of solid-state fermentation products,addition of Mn could increase the low polarity metabelites, but has little effect on the mediate and high polarity metabelites. Addition of 1 ppm of Ge, Zn and 7.5 ppm of Mn showed similar results in the fermentation products with herbal medicines(HAC). The polysaccharide contentswere increased with Fe addition, while the addition of Se, Ge and low concentration of Mn, Zn enhanced the total phenol contents. On the other hand, low concentration of Mn, Zn improved the DPPH scavenging ability. Addition of Se, Ge and Mn improved the chelating ability of AC mycelia on ferrous ion. For the HAC, addition of microelements possessed little effect on the polysaccharide content, while the total phenol content and ABTS scavenging abilities could by improved by the addition of Ge and low concentration of Se, Fe and Zn. The DPPH scavenging abilities could be incressed with addition of Ge and Mn, while the ability of chelating ferrous ion could be enhanced by the addition of Mn, Fe and low concentration of Ge. As for the in vitro tumor cell tests, addition of microelements could enhanced the suppressibility of fermentation products on the growth of HCT-8 and Hep G2 tumor cell.
In conclusion, the addition of suitable microelements with proper dosage on the AC fermentation could enhance the bioactivities of the products in many aspects.
第一章 緒論··················································································································1
1.1 研究背景··········································································································1
1.2 研究動機與目的······························································································2
第二章 文獻回顧··········································································································3
2.1 牛樟芝文獻回顧····························································································3
2.1.1 牛樟芝介紹···························································································3
2.1.2 牛樟芝成份及生理活性試驗·······························································4
2.1.3 牛樟芝之藥理活性···············································································8
2.2 氧化與抗氧化關係·······················································································10
2.2.1 脂質氧化作用······················································································10
2.2.2 抗氧化劑······························································································11
2.3 微量元素文獻回顧·······················································································12
2.3.1 鋅的研究發展與生物學功能······························································12
2.3.2 鐵的研究發展與生物學功能······························································13
2.3.3 鍺的研究發展與生物學功能······························································14
2.3.4 錳的研究發展與生物學功能······························································15
2.3.5 硒的研究發展與生物學功能······························································16
2.4 中草藥文獻回顧··························································································17
2.4.1 白花蛇什草·························································································17
2.4.2 決明子·································································································18
2.4.3 板藍根·································································································19
2.4.4 魚腥草·································································································20
2.4.5 枸杞子·································································································21
2.4.6 三七·····································································································22
第三章 實驗架構········································································································24
第四章 實驗材料與方法····························································································25
4.1 牛樟芝菌種與中草藥配方··········································································25
4.2 實驗儀器設備·····························································································25
4.3 實驗藥品·····································································································26
4.4 實驗方法·····································································································27
4.4.1菌種保存······························································································27
4.4.2 平盤培養·····························································································27
4.4.3 液態發酵培養·····················································································28
4.4.4固態發酵培養······················································································29
4.4.5 測試微量元素對牛樟芝發酵產物之生理活性之測定·····················29
4.4.6 測試微量元素對牛樟芝發酵產物多醣含量之測定·························30
4.4.7測試微量元素對牛樟芝發酵產物抗氧化物質含量測定··················33
4.4.8 測試微量元素對牛樟芝發酵產物體外抗氧化活性之影響·············35
4.4.9 測試微量元素培養下其發酵產物對腫瘤細胞生長之影響·············39
4.5 統計分析·····································································································40
第五章 結果與討論····································································································41
5.1 微量元素對牛樟芝液態發酵菌絲體生長之影響······································41
5.2 微量元素對牛樟芝固態發酵物之成份分析··············································44
5.3 微量元素對牛樟芝液態發酵物之多醣產量影響·····································48
5.3.1 牛樟芝液態發酵液之多醣產量影響················································48
5.3.2 牛樟芝液態菌絲體之多醣產量影響················································50
5.4 微量元素對牛樟芝固態發酵物之多醣產量影響······································51
5.5 微量元素對牛樟芝液態發酵物之抗氧化能力影響··································53
5.5.1 牛樟芝液態發酵液之抗氧化能力影響············································53
5.5.2 牛樟芝液態菌絲體之抗氧化能力影響············································60
5.6 微量元素對牛樟芝固態發酵物之抗氧化能力影響··································64
5.7 微量元素對牛樟芝液態發酵產物抑制腫瘤細胞生長活性之影響··········71
5.7.1 對牛樟芝液態發酵液抑制腫瘤細胞生長活性之影響·····················71
5.7.2 對牛樟芝液態菌絲體抑制腫瘤細胞生長活性之影響·····················79
5.8 微量元素對牛樟芝固態發酵產物抑制腫瘤細胞生長活性之影響··········83
第六章 結論················································································································91
第七章 參考文獻········································································································93
附件一·························································································································105
附件二·························································································································106
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