| 摘要: | 紅景天是中國傳統藥用植物之一，其主要藥用部位是根和莖。根據文
獻報導記載，紅景天具有抗缺氧、抗疲勞、抗腫瘤、抗病毒、抗老化、抗
氧化及抗輻射等多種幼纂C然而，目前紅景天於化粧品用途應用上並不多
見，尤其在化粧品美白及抗紫外線傷害部份的開發及應用更是缺乏相關研
究實驗探討。因此，本論文研究主題便是針對紅景天於美白及抗紫外線傷
害幼臚W的評估為主，並試圖找出其可能的作用機制。此方面相關實驗進
行主要分為兩部分來探討，分述如下。
第一部份實驗著重於美白幼蘆熊禲A利用黑色素瘤細胞(B16 cells)經
紅景天水或甲醇萃取液處理後，觀察對黑色素生合成與酪胺酸酶活性之影
響。我們利用酵素連結免疫吸附法(Enzyme-Linked ImmunoSorbent Assay;
ELISA)，分別來檢測兩種紅景天萃取液對體外酪胺酸酶(tyrosinase)抑制率
及B16 細胞內酪胺酸酶活性的影響。此外，以細胞存活率測試(MTT assay)
來觀察兩種紅景天萃取液對B16 細胞生長速率的影響，並進一步分析B16
細胞黑色素含量的變化，以確定紅景天萃取液對黑色素生成之影響。結果
顯示，5 mg/ml 紅景天水萃取液(water extracts of Rhodiola; WER)或0.1 mg/ml
甲醇萃取液(methanol extracts of Rhodiola; MER)對試管酪胺酸酶活性測試
有較佳的抑制效果，抑制率分別為
35 %和79 %。0.5-5 mg/ml WER 或0.01-0.1 mg/ml MER 在無影響B16 細胞
生長情況經24 小時培養後，其細胞內的酪胺酸酶活性與黑色素生成量明顯
的被抑制且呈現劑量關係。5 mg/ml WER 與1 mg/ml MER 對細胞內的酪胺
酸酶抑制率分別為13 %和12 %，而對B16 細胞內的黑色素生成量的抑制
率也各別為68 %和67 %。
II
第二部份實驗主要是針對抗紫外線傷害幼蘆熊禲A利用受到紫外線
(UV 365 nm, 8 J/cm2)照射的人類纖維母細胞(Hs68 cells)，經WER 或MER
前處理後，以彗星分析法觀察對紫外線誘發細胞DNA 損傷之影響。同時也
監測有無WER 或MER 處理，經誘發細胞DNA 損傷後其細胞內脂質過氧
化變化情形。此外我們也進一步完整評估WER 和MER 的抗氧化能力程度
(總抗氧化力、抑制過氧化率、清除超氧陰離子測定、清除DPPH 自由基的
能力及蟄合鐵離子效應檢測)。結果顯示，處理0.5-5 mg/ml WER 或0.01-0.1
mg/ml MER 經24 小時培養後，以UV 造成Hs68 細胞DNA 損害的情形，
明顯獲得改善，並呈現劑量關係。對UV 誘導細胞DNA 傷害，經5 mg/ml
WER 和1 mg/ml MER 前處理後，其tail moment 值分別從14 降低到2 和15
下降至4。此外，對於抗氧化能力程度的評估，經WER 或MER 處理後，
程度皆顯著提升。
根據上述實驗結果得知，紅景天美白的幼艦i能經由抑制酪胺酸酶活
性而使黑色素含量降低。而針對抗紫外線傷害的幼蘋h可能藉由提升抗氧
化能力程度而減輕因UV 誘導的細胞DNA 傷害情形。因此，我們推論紅景
天於化粧品美白及抗紫外線傷害部份的開發及應用深具潛力，值得深究。
Rhodiola (Rhodiola rosea L.) is kinds of Chinese traditional herbs. The roots
and stems of Rhodiola are mainly the medicinal region. According to reports
that the Rhodiola had various efficacies, such as in the aspects of anti-hypoxia,
anti-fatigue, anti-cancer, anti-bacteria, anti-aging, anti-oxidation and
anti-irridation. However, there was less attention on application in cosmetics. It
lacks for related investigations especially about cosmetic whitening and
anti-ultraviolet (UV)-induced damage in development and application of
Rhodiola. Hence, the studying subjects of this thesis are mainly focused on
evaluations of whitening and anti-UV-induced damage efficiency, and trying to
find the probably underlying mechanisms out. The experimental processes of
this study are divided into two parts. It describes as following.
The first part of this study is emphasized on estimation of the whitening
efficacy. We observed that the effects of treatment with water extracts of
Rhodiola (WER) or methanol extracts of Rhodiola (MER) in melanoma B16
cells on melanogenesis and tyrosinase activity. The influences of tyrosinase
inhibitiory percentage in vitro and in B16 cells with WER or MER were
respectively measured by using the method of Enzyme-Linked ImmunoSorbent
assay (ELISA). Additionally, the effects of WER or MER on the growth of B16
cells were also evaluated by using the method of MTT assay, and analyzed the
changes of melanin content. Our results showed that treatment of 5 mg/ml WER
or 0.1 mg/ml MER had better outcomes in tyrosinase inhibitory rate, and the
values were 35 % and 79 %, respectively. The intracellular tyrosinase activity
and melanin content were significantly suppressed and played the correlation in
IV
a dose manner, under no abnormal growth changing condition after 24 hr
incubation in B16 cells with 0.5-5 mg/ml WER or 0.01-0.1 mg/ml MER. The
values of tyrosinase activity inhibitory rate in B16 cells with 5 mg/ml WER and
1 mg/ml MER were 13 % and 12 %, and the values of melanogenesis inhibitory
rate were 68 % and 67 %, respectively.
The second part of this study is concentrated on assessment of anti-
UV-induced damage efficacy. We examined that the effects of pretreatment with
WER or MER in ultra-violet (UV 365 nm, 8 J/cm2)–irradiated human fiberoblast
cells (Hs68 cells) on situation of cell DNA damage by the method of comet
assay. Meanwhile, the lipid peroxidation was also detected with or without WER
or MER pretreatment during UV irradiation-induced DNA damage in Hs68 cells.
Furthermore, the degree of antioxdative capacity on WER or MER were also
estimated via trolox equivalent antioxidant capacity, inhibition of peroxidation
effects, superoxide anion radical scavenging effects, DPPH radical scavenging
effects, and chelating action on ferrous ion effects. Results revealed that the
circumstances of UV-induced DNA damage were apparently ameliorated and
played the correlation in a dose manner after 24 hr incubation in Hs68 cells with
0.5-5 mg/ml WER or 0.01-0.1 mg/ml MER. The values of tail moment after
pretreatment of 5 mg/ml WER and 1 mg/ml MER were respectively reduced
from 14 to 2 and from 15 to 4 in the UV-indcued Hs68 DNA damage. Beside
that, the levels of antioxidative capacity with administration of WER or MER
were all remarkably raised than other vehicle treatment.
The present results implied that the whitening function of Rhodiola extracts
might be diminished the melanin content through tyrosinase activity inhibition,
V
and the function of anti-UV-induced damage might be mediated by enhancing
antioxidant capacity to prevent the cell DNA damage. Therefore, we speculate
that Rhodiola extracts may be used as the valid and safe whitening and anti-UV
damage composition of cosmetics. It seems to have great potential to develop
the application of Rhodiola in cosmetics. |
