Study on Anti-oxidation, Anti-inflammatory, Whitening and Skin Repair by Means of Callus Induction and Cell Suspension Culture of Plants

• Main Authors: Zih-Ling Wu, 伍姿綾
• Other Authors: Jane-Yii Wu
• Format: Others
• Language: zh-TW
• Published: 2016
• Online Access: http://ndltd.ncl.edu.tw/handle/s2s7t5

碩士 === 大葉大學 === 生物產業科技學系 === 104 === Aquilaria sinensis is widely used in herbal medicine and containing many high-value compounds. However, these compounds are difficult to cultivate or are becoming endangered because of overharvesting. Therefore, an in vitro culture of Aquilaria sinensis was established for production of commercially important secondary metabolites. Callus cultures were obtained by inoculating leaf explants on Murashige and Skoog (MS) medium supplemented with 6-BA alone or in combination with either α-naphthalene acetic acid or 2,4-D. The callus obtained in response to 2,4-D and 6-BA was subcultured on the same mediumto investigate its biomass accumulation, and callus production on weekly basis for 43 days. For submerged cultivation, old calli were cultured on MS basal media supplemented with 2,4-D and 6-BA. Additionally, Fourier transform infrared spectroscopy(FTIR) was carried out in the water/ethanol extracts from both the wild plants and cell suspension cultures of Aquilaria sinensis. On the other hand, to evaluate the level of the anti-oxidation, anti-inflammatory, and skin repair activities of the water/ethanol extracts from both the wild plants and cell suspension cultures of Aquilaria sinensis. The results indicate that WEASL and EEASL sample have highest antioxidant activity. The anti-inflammatory activity of the water/ethanol extracts from both the wild plants and cell suspension cultures of Aquilaria sinensis in lipopolysaccharide-stimulated murine RAW 264.7 cells was evaluated. The present study suggested that purified component, the water/ethanol extracts from both the wild plants and cell suspension had strong anti-inflammatory effects on LPS-stimulated RAW264.7 macrophages through inhibiting NF-κB secretion levels reduced COX-2, iNOS and that NO and cytokines release. This study is to investigate the influence of the water/ethanol extracts from both the wild plants and cell suspension on the wound healing. The ethanol extracts from cell suspension was shown to significantly promote the migration of fibroblasts in wound scratching assay. The superior effect of the ethanol extracts from cell suspension was observed at 1mg/mL, in which the recover rates was increased with 2 and 3 folds after 12 h and 24 h respectively than that of blank control (P < 0.05). UV radiation is known to induce free radical formation in irradiated tissues such as skin, in a process involving lipid peroxidation. The skin repair activity of the ethanol extracts from cell suspension cultures of Aquilaria sinensis in UVB-stimulated murine CCD-966ks cells was evaluated. In the in vitro wound healing assay, the ethanol extracts from cell suspension, at 12 h and 24 h, enhanced fibroblasts migration. Aquilaria sinensis leaf production in cell culture can be modulated, and more importantly, secondary metabolites accumulation could be increased. All the tested extracts of the water/ethanol extracts from both the wild plants and cell suspension could exhibit the anti-oxidation, anti-inflammatory and skin repair activities to some extents. These results the water/ethanol extracts from both the wild plants and cell suspension were shown a promising natural drug with anti-oxidation, anti-inflammatory and skin repair effects for wound treatment.