牛樟芝(Antrodia cinnamomea)為台灣本土特有種的藥用真菌，根據研究指出其子實體或菌絲體具有保護肝臟、抗氧化、抗發炎及抑制腫瘤等功效。故本研究藉牛樟芝菌絲體液態深層培養來醱酵5種中草藥，乃是利用牛樟芝菌絲體的酵素系統對中草藥的成份進行生物轉化作用(bio-transformation)；另外，進一步分析牛樟芝醱酵各種中草藥後產生的轉化代謝物(bio-transformed metabolites)對於皮膚抗老化之活性。研究結果顯示，處理濃度為5.0%時，只有牛樟芝丹參醱酵液具細胞毒性，其餘醱酵液之細胞存活率皆在80.0%以上；而當處理濃度提高至10.0%時，則牛樟芝桑白皮及女貞子之醱酵液具有較低的細胞毒性，其餘醱酵液之細胞存活率甚低。另一方面，在酵素活性抑制分析試驗中，以最終濃度5.0%的牛樟芝丹參、女貞子及牡丹皮之醱酵液為較具抑制MMP-2及MMP-9的作用，當濃度提高至10.0%時抑制效果越趨顯著。再從5種中草藥醱酵液中挑選出丹參、女貞子及牡丹皮進行膠原蛋白保護之含量分析，發現添加了中草藥的醱酵液可達到延緩膠原蛋白破壞之效果，值得注意的是牛樟芝丹參醱酵液保護膠原蛋白被降解的能力為最強。因此，綜合上述實驗結果，我們認為利用牛樟芝醱酵中草藥萃取物之生物轉化代謝物的確具有可探討性，並可應用於抗老化化粧品原料之開發。

Antrodia cinnamomea is an endemic fungus grown in Taiwan. Recent studies demonstrated that the fruiting body or mycelia of A. cinnamomea have several bioactivities, including liver protection, anti-oxidation, anti-inflammation and anti-tumor. In this study, A. cinnamomea mycelial submerged culture is applied to perform the fermentation with 5 medicinal herbal extracts. The enzymes presented in mycelial submerged culture of A. cinnamomea will be employed to carry out bio-transformation of medicinal herbal extracts. Additionally, the objective of this project is planned to investigate the antiaging activities of bio-transformed metabolites isolated from A. cinnamomea submerged culture which co-fermented with those medicinal herbal extracts. The experimental results showed that when 3T3 cell were treated with the concentration of 5.0%, cytotoxicity was found only from the A. cinnamomea submerged fermentative Salvia miltiorrhiza product; as well as, the cell survival rates of other fermented A. cinnamomea medicinal herbal extracts are above 80.0%. When treated with the concentration which increased to 10.0%, the products from A. cinnamomea submerged fermentation with Morus alba, and Ligustrum lucidum showed low cytotoxicities; however, survival rates of the others was relatively low. On the other hand, the analyses by gelatin based zymography showed that the products from A. cinnamomea submerged fermentative Salvia miltiorrhiza, Ligustrum lucidum, and Paeonia suffruticosa, exhibited the abilities of inhibition toward MMP-2 and MMP-9 with the concentration of 5%; moreover, inhibitive capacities were remarkably increased when the concentration elevated 10.0%. As a result, we selected the Salvia miltiorrhiza, Ligustrum lucidum, and Paeonia suffruticosa fermentative products from 5 kinds of medicine herbal fermentative products proceeded to collagen protection assay. We found that collagen degradation was delayed by supplementation of medicinal herbal extracts fermentative products. It is worth noticing that A. cinnamomea submerged fermentative Salvia miltiorrhiza product showed the strongest ability to promote collagen accumulation. In conclusion, we suggest that future researches can confer with bio-transformation of medicinal herbal extracts by A. cinnamomea, and these bio-transformed products can be developed as effective antiaging agents used in skincare cosmetics.

中文摘要 I
表目錄 VII
圖目錄 VIII
縮寫表 XI
第一章 緒論 1
1-1. 前言 1
1-2. 研究動機與目的 3
1-3. 研究架構 4
第二章 文獻回顧 5
2-1. 人體皮膚構造 5
2-2. 皮膚與老化 9
2-3. 膠原蛋白 11
2-4. 基質金屬蛋白酶 15
2-5. 牛樟芝簡介 20
2-5-1. 牛樟芝分類與特徵 21
2-5-2. 牛樟芝的生理活性成分及其功效 22
2-6. 中草藥文獻回顧 24
2-6-1. 黃耆Astragalus membranaceus 24
2-6-2. 女貞子Ligustrum lucidum 24
2-6-3. 桑白皮Morus alba 25
2-6-4. 牡丹皮Paeonia suffruticosa 25
2-6-5. 丹參Salvia miltiorrhiza 26
第三章 研究材料、儀器與方法 27
3-1. 研究材料 27
3-2. 實驗方法 31
3-2-1. 中草藥之水萃取法 31
3-2-2. 牛樟芝菌株培養 31
3-2-3. 細胞活化與繼代培養 33
3-2-4. 體外動物細胞存活率分析 34
3-2-5. 基質金屬蛋白酶活性抑制分析 35
3-2-6. 膠原蛋白濃度分析 36
第四章 結果與討論 38
4-1. 不同中草藥萃取物添加於A. cinnamomea固態培養基及液態醱酵培養其菌絲體之影響 38
4-1-1. 不同中草藥萃取物添加於A. cinnamomea固態培養基其菌絲體生長型態之觀察 38
4-1-2. 添加不同中草藥萃取物對於A. cinnamomea固態培養基之菌絲體生長之影響 46
4-1-3. 不同中草藥萃取物添加於A. cinnamomea液態醱酵培養其菌絲體之觀察 48
4-2. 以A. cinnamomea中草藥醱酵液處理3T3纖維母細胞之細胞存活率分析 56
4-3. A. cinnamomea中草藥醱酵液之MMP-2及MMP-9活性抑制分析 70
第五章 結論 83
參考文獻 85
 

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