| 摘要: | (PART1)目前化粧品仍以人工製造之化學原料為主，這些原料雖無明確研究指出其對環境所造成的衝擊，但其實際上不具備永續利用的理念，對於健康所可能產生的風險亦是未知。因此在化粧品中添加天然萃取物已成為現今趨勢，本研究以常作為行道樹種植的植物莖部酒萃物(ZLSE)、植物莖部水萃物(ZLSW)、植物葉酒萃物(ZLLE)、植物葉水萃物(ZLLW)進行抗氧化、美白、抗發炎、預防光老化及DNA修復之機制探討。
結果顯示ZLSE、ZLSW、ZLLE和ZLLW具有較良好清除DPPH·和ABTS·+自由基能力，在還原力與螯合亞鐵試驗亦有顯著效能，而在脂質過氧化試驗中ZLSE、ZLSW、ZLLE和ZLLW與對照組Trolox的效果相當甚至部分超越。然而在總酚、總黃酮、總花青素及總多醣含量試驗中ZLSE、ZLSW、ZLLE和ZLLW並無明顯差異，表示皆具備良好的抗氧化潛力。並且在抑制酪胺酸酶試驗中發現ZLSE、ZLLE和ZLLW有促進黑色素生成的趨勢。
在進一步的細胞內試驗的部分，由於ZLSE及ZLLE在細胞存活度試驗中顯示其具有毒性，因而後續實驗以ZLSW和ZLLW進行。在抗氧化試驗中ZLSW和ZLLW可有效抑制HaCaT細胞內活性氧物質(reactive oxygen species, ROS)與超氧陰離子(superoxide, O2.-)生成，並提高穀胱甘肽(reduced glutathione, GSH)含量；酵素性抗氧化系統超氧歧化酶(superoxide dismutase, SOD)、榖胱甘肽過氧化酶(glutathione peroxidase, GPx)和過氧化氫酶(catalase)之表現亦有上升趨勢。美白試驗則利用B16F10細胞進行，結果顯示ZLSW與ZLLW具有提升黑色素生成之活性，且提高黑色素生合成相關蛋白質轉錄因子(melanocyte inducing transcription factor, MITF)、melanocortin-1 receptor (MC1R)、酪胺酸酶(tyrosinase)、和TRP-1 (tyrosinase-related protein-1)表現。抗發炎效能評估同樣利用HaCaT細胞進行試驗，發現ZLSW、ZLLW均具降低發炎相關因子腫瘤壞死因子-α (tumor necrosis factor-α, TNF-α)、介白素1β (interleukin-1β, IL-1β)和介白素6 (IL-6)的表現，並使mitogen-activated protein kinase (MAPK)家族的活化降低。預防光老化試驗透過3T3L1細胞證實ZLSW、ZLLW具降低基質金屬蛋白酶(matrix metalloproteinase, MMPs)生成及促進膠原蛋白生成的能力。利用HaCaT細胞證實ZLSW和ZLLW能降低經UVB誘發所產生之光化產物環丁烷嘧啶二聚體(cyclobutane pyrimidine dimers, CPDs)和6-4二聚體(6-4 photoproducts, 6-4PPs)，進而促使DNA修復。且ZLSW、ZLLW是藉由調控核苷酸切除修復機制(nucleotide excision repair, NER)相關蛋白質，來修復經UVB誘發所造成的DNA損傷。
由上述試驗可知，ZL水萃物具有抗氧化、抗發炎、抗光老化、修復及促進黑色素生成之功效，未來若能應用於化妝品原料開發上將是一具備良好潛力的材料，且由於其促進黑色素生成之效果更加適合應用於毛髮相關商品上。
(PART2)頭頸部癌(head and neck squamous carcinoma, HNSCC)泛指發生在鼻咽、口腔、喉、及唾液腺等部位的癌症，其發生率很高，乃是國人十大癌症中死亡率排名第五的癌症。目前針對口腔癌的臨床治療方式仍以手術開刀切除腫瘤並配合全身性化學治療和放射線治療為主，理論上若是以手術完全移除腫瘤細胞，癌症是可以被治癒的，但外科手術通常無法切除已經轉移到其他部位的癌細胞，導致頭頸部癌患者預後程度不佳。
本研究利用IL-S RP及siRNA對台灣本土口腔癌細胞(keratinocytes of a human gingival squamous carcinoma cell, OECM-1)進行作用，探討IL-S過度表現或缺失對癌細胞之生長與其對上皮間質轉化(epithelial-mesenchymal transition, EMT)路徑之影響。實驗證實IL-S RP在200 ng/ml的濃度作用下，具促進OECM-1細胞生長的作用，而IL-S siRNA反之。並且IL-S RP在200 ng/ml的濃度作用24小時後明顯促進OECM-1發生遷移，而EMT是細胞遷移的重要過程，故進一步利用即時聚合酶鏈鎖反應及西方墨點法證實IL-S透過interleukin-6 (IL-6)、epidermal growth factor receptor (EGFR)及signal transducer and activator of transcription 3 (STAT3)與mitogen protein kinase (p38)、extracellular regulated protein kinase (ERK)、B-cell lymphoma 2 (Bcl-2)及hypoxia-inducible factor-1-alpha (HIF-1)等兩條路徑活化vimentin及fibronectin等EMT相關因子，並同時降低E-cadherin表現促使EMT發生。在不同分期病人組織內同樣觀察到與上述相同的路徑，並且此影響只針對腫瘤組織，顯示IL-S促進EMT發生的路徑具有對頭頸部癌的專一性。而在正常人與頭頸部癌患者的血清中亦發現IL-S的含量明顯於頭頸部癌患者體內普遍較高，未來可望透過IL-S提早發現頭頸部癌發生的風險，並加以預防、治療。
Nowadays, cosmetics are mainly made of artificial chemical materials. Although there is no clear research indicates the impact of these materials in environment, they actually do not have the concept of sustainable use, and the dangers that may be caused to health are also unknown. Therefore, it becomes a trend to add natural extracts in cosmetics.
In this study, we chose ZL which are often plants as sidewalk trees to investigate the mechanism of anti oxidation, whitening, anti inflammation, prevention of photoaging and DNA repair by ZLSE, ZLSW,ZLLE, and ZLLW, respectively.
Results show that the ZLSE, ZLSW, ZLLE and ZLLW have great ability of DPPH. and ABTS.+ free radical scavenging. In the tests of reducing power and ferrous iron chelating have significant efficacy. In the lipid peroxidation test, the effects of ZLSE, ZLSW, ZLLE and ZLLW were similar to or even exceeded by Trolox.
However, there was no significant difference between ZLSE, ZLSW, ZLLE and ZLLW in total phenols, total flavonoids, total anthocyanins and total polysaccharides which means all of the above have great ability(potentiality) of an anti oxidation. In addition, it was found that ZLSE, ZLLE and ZLLW had a tendency to promote melanin production in the experiments of inhibiting tyrosinase.
Moreover, since ZLSE and ZLLE were shown to be toxic in cell viability assays, subsequent experiments were performed with ZLSW and ZLLW. In the intracellular antioxidant assay, ZLSW and ZLLW could effectively inhibit the production of reactive oxygen species (ROS) and superoxide (O2.--) in HaCaT cells, and increased the content of glutathione
(GSH). The performance of superoxide dismutase (SOD), glutathione perxidase (GPx) and catalase also showed an increasing trend. Whitening test was conducted on B16F10 cells, and the results showed that ZLSW and ZLLW could enhance melanin production, and improve the performance of melanocyte inducing transcription factor (MITF), melanocortin--1 receptor (MC1R), tyrosinase, and tyrosinase--related protein--1 (TRP--1).
HaCaT cells were used to evaluate the anti--inflammatory efficacy, and it was found that ZLSW and ZLLW could both decrease the performance of tumor necrosis factor--α (TNF--α), interleukin--1β (IL--1β) and IL--6, and reduced the activation of the mitogen--activated protein kinase (MAPK) family.
The photoaging prevention tests confirmed that ZLSW and ZLLW had the ability of reducing matrix metalloproteinase (MMPs) and increase collagen synthesis by 3T3L1 cells. HaCaT cells were used to confirm that ZLSW and ZLLW could reduce the photochemical products induced by UVB irradiation, such as cyclobutane pyrimidine dimers (CPDs) and 6--4photoproducts (6--4PPs), and thus promoted DNA repair. Moreover, ZLSW and ZLLW could repair DNA damage induced by UVB through regulating relevant proteins of nucleotide excision repair (NER) system.
According to the present study, ZL water extracts has the effect on anti--oxidation, anti--inflammation, anti--photoaging, repair and induce melanin production. ZL water extracts will be a promising material in the future if it can be applied to the development of cosmetic raw materials, and it is more suitable for the hair related commodities in particular because of its performance on inducing melanin production. |
