| 摘要: | 本研究主要的目的是將常見之蔬菜廢棄物加以利用，應用於化粧品原料開發，提升其經濟價值。在市場中挑選十六種常見被丟棄之蔬菜部位，包括白蘿蔔(Raphanus sativus)葉、皇帝豆(Phaseolus lunatus)殼、小白菜(Brassica rapa chinensis)葉、芹菜(Apium graveolens)葉、花椰菜(Brassica oleracea L. var. botrytis L.)梗部、蘆筍(Asparagus officinalis)皮、茭白筍(Zizania latifolia)殼、茼蒿(Chrysanthemum coronarium)葉、結球白菜(Brassica rapa，俗稱大白菜)葉、結球甘藍(Brassica oleraceae L. var. capitata L.，俗稱高麗菜)葉、青江菜(Brassica chinensis)葉、水菜(Brassica juncea)葉、黑葉白菜(Brassica campestris)葉、翠白菜(Brassica pekinensis)葉、竹筍(Phyllostachys edulis)殼及桑(Morus alba)葉。
將蔬菜的不同部位以水洗淨、瀝乾，以果汁機撹碎後進行水萃取，經過濾去除殘渣，將過濾後之萃取液分別進行以Nitrobule tetrazolium (NBT)為受質之超氧歧化酶活性測試及超氧歧化酶抑制自由基之測定，爾後挑選出酵素活性較強之高麗菜葉萃取液(BOCE)、花椰菜梗部萃取液(BOBE)及桑葉萃取液(MAE)進行體外細胞毒性測試(MTT assay)，並將樣品分別以不同倍數稀釋，應用於抑制3T3纖維母細胞所分泌之基質金屬蛋白酶(matrix metalloproteinases, MMPs)，藉由gelatin-based zymography進行MMP-2和MMP-9之活性抑制分析，測試其是否具有抑制MMPs活性的能力。另外，透過抑制2', 7'-dichlorofluorescin-diacetate(DCFH)氧化相關分析試驗，進一步對植物萃取液中非酵素性抗氧化能力進行分析評估。
超氧歧化酶活性染色之實驗結果顯示，花椰菜梗部萃取液(BOBE)、高麗菜葉萃取液(BOCE)及桑葉萃取液(MAE)具有較明顯的活性；而在超氧歧化酶抑制自由基的比率也都超過96%。
在細胞毒性分析方面，除了桑葉萃取液在高濃度時會使細胞存活率降至60%~70%外，其餘的均無明顯之影響。
另一方面，花椰菜梗部及高麗菜葉之萃取液對於基質金屬蛋白酶活性並沒有明顯的抑制作用，但桑葉萃取液在處理48小時期間內有抑制基質金屬蛋白酶MMP-9的活性。而高麗菜葉萃取液(BOCE)及桑葉萃取液(MAE)之非酵素性抗氧化能力分別在6.62 mg/ml及6.27 mg/ml之濃度下均有70%~80%之抑制率。
This research aims to investigate whether the vegetable extracts contain high activities of enzymatic antioxidants (superoxide dismutase, SOD) and assess the potentials of these extracts applied in cosmetic industry.
Sixteen common vegetable wastes were collected for activity assay, including Raphanus sativus leaf , Phaseolus lunatus shell, Brassica rapa chinensis leaf, Apium graveolens leaf, Brassica oleracea L. var. botrytis L. stalks, Brassica juncea leaf, Asparagus officinalis peel, Zizania latifolia shell, Chrysanthemum coronarium leaf, Brassica rapa leaf, Brassica oleraceae L. var. capitata L. leaf, Brassica chinensis leaf, Brassica campestris leaf, Brassica pekinensis leaf, Phyllostachys edulis shell, and Morus alba leaf. All vegetable wastes were water-extracted and filtered to harvest extracts, respectively.
Extracts were then examined by SOD activity staining analysis via a series of NBT verification proceduces, cell viability assay with a tetrazolium dye, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), gelatin-based zymography, and a cell-based assay, using 2', 7'-dichlorofluorescin (DCFH) oxidation.
To begin with, SOD activity staining analysis revealed that three vegetable extracts [ i.e. the extract of Brassica oleracea L. var. botrytis L. stalks (BOBE), the extract of Brassica oleraceae L. var. capitata L. leaves (BOCE), and the extract of Morus alba leaves (MAE) ] were of significant activities and the rate for inhibiting free radicals of SOD exceeded 96% .
Secondly, the MTT assay confirmed that high concentrations of MAE declined the cell survival rate to 60-70%, but such declination did not apply to other vegetable extracts aforementioned.
Thirdly, BOBE and BOCE did not inhibit the activities of MMP-2 and MMP-9, whereas MAE showed significant MMP-9 inhibition during 48 hours treatment.
Moreover, the activities of non-enzymatic antioxidants in BOCE and MAE exhibited 70~80% of inhibition in DCFH oxidation at 6.62 mg/ml and 6.27 mg/ml, respectively. |
