| 摘要: | 桑椹為桑科植物 Morus alba L. 的乾燥果穗，其性狀呈黃棕色、棕紅色或暗紫色，功能為滋陰補血，而臺灣中藥典無桑椹之品質規範。桑椹富含多酚類和多醣類，具有抗氧化與抗發炎生物活性。因此以性狀、化學分析、抗氧化及抗發炎活性分析，探討桑椹其生物品質管制指標。收集不同來源的桑椹，利用 L*a*b* 色差值測量 30 件市售乾燥桑椹，並結合主成分分析，發現10件 L*<35、*a<1、*b<8 為紫色樣品，與 20 件 L*>35、*a>1、*b>17 為黃色樣品。繼而分析樣品中的總多酚、總黃酮、總花青素、糖度、pH值、Chlorogenic acid、Cyanidin-3-glucoside （C3G）、Cyanidin-3-Rutinoside（C3R）、Rutin、Quercetin等指標成分。結果經層次聚類熱圖分群顯示總多酚、總黃酮、總花青素、C3G、C3R、糖度、pH值在紫桑椹較多，而總多醣、Quercetin、Rutin、Chlorogenic acid則在黃桑椹較多。而不同生長期樣品可分為未成熟期 (ST1)、初熟期 (ST2)、成熟期 (ST3) 及老化期 (ST4) ，發現Chlorogenic acid於ST2含量分佈中位數最高、ST4 最低。C3G及C3R含量隨成熟度增加而上升於ST4達到最高。Rutin、Quercetin含量在不同生長階段變化無顯著差異。據此結果，推測黃桑椹屬ST2，而紫桑椹屬ST4。將30件市售品進行DPPH及ABTS自由基清除試驗、H2O2誘導BV2損傷試驗、LPS誘導RAW264.7及BV2產生NO試驗。結果經由層次聚類熱圖分群顯示30件市售品可分為三大群，抗氧化組主要為紫桑椹，而抗發炎組及混合組主要為黃桑椹，再將三組進行活性與成分相關性分析，發現混合組活性與 Quercetin含量正相關，建議Quercetin可為黃桑椹的生物品質管制指標。抗氧化組活性與總多酚、總黃酮、總花青素、C3G、C3R 正相關。市售樣品可測得C3G及C3R樣品比例低（40%），且總花青素含量高易檢測，故建議以總花青素為紫桑椹之生物品質管制指標。繼而利用F16及F17樣品探討紫與黃桑椹的活性差異。結果顯示，F17 比F16 更顯著抑制LPS 誘導BV2 細胞突觸縮短、iNOS 及COX-2 之蛋白質表現，具有較強的抗發炎作用。反之，於H2O2誘導 BV2 細胞的氧化損傷模式，F16比F17更顯著活化HO-1蛋白質，具有較強的抗氧化作用。
Mulberry is the fruit of Morus alba L., a member of the Moraceae family. The fruit color varies from yellowish-brown, brownish-red, to dark purple. It is traditionally used to nourish yin and blood. However, the pharmacopeia still has no established standards to classify the fruit according to physical properties and chemical composition. Mulberry are rich in polyphenols and polysaccharides known for their antioxidant and anti-inflammatory capacity. Therefore, this study aims to investigate the properties and Q-biomarker of mulberry through characterization, chemical analysis, and the evaluation of its antioxidant and anti-inflammatory properties. Mulberries were collected from various sources. Thirty commercially available dried mulberries were collected from different sources and evaluated by using L*a*b* color and then for PCA analysis. 10 samples with L*<35, a*<1, b*<8 were purple, while 20 samples with L*>35, a*>1, b*>17 were yellow. Moreover, the chemical analysis included total polyphenols, total flavonoids, total anthocyanins, sugar, pH, chlorogenic acid, cyanidin-3-glucoside (C3G), cyanidin-3-rutinoside (C3R), rutin, and quercetin as major components. The HCA cluster results indicated that purple samples had higher levels of total polyphenols, total flavonoids, total anthocyanins, C3G, C3R, sugar, and pH, while yellow samples had higher levels of total polysaccharides, quercetin, rutin, and chlorogenic acid. Samples from different growth periods were categorized into four stages: immature stage (ST1), early ripening stage (ST2), ripening stage (ST3), and aging stage (ST4). Chlorogenic acid was found to be the highest in the median of ST2 and the lowest in ST4, while the contents of C3G and C3R increased with the increase of ripening and reached the highest in ST4. There was no significant difference in the contents of Rutin and Quercetin in different growth stages. Based on these results, it was hypothesized that yellow mulberry belongs to ST2 and purple mulberry belongs to ST4.The thirty samples underwent DPPH and ABTS free radical scavenging tests, H2O2-induced BV2 damage tests, and LPS-induced RAW264.7 and BV2 NO production tests. HCA results of bioactivities showed that the 30 market samples could be classified into three groups. The antioxidant group was predominantly purple, while the anti-inflammatory and mixed groups mainly consisted of yellow samples. Activities and components correlation analysis for each group revealed that the activity of the mixed group was positively correlated with quercetin, which was suggested to be a Q-biomarker of yellow mulberry.The activity of the antioxidant group was positively correlated with total polyphenols, total flavonoids, total anthocyanins, C3G and C3R. The proportion of C3G and C3R in the commercially available samples was low (40%) , and the content of total anthocyanins was high and easy to detect, therefore, it was suggested that total anthocyanins should be used as the Q-biomarker of purple mulberry. Finally, F16 and F17 samples were used to investigate the activity differences between purple and yellow mulberry. The results showed that F17 inhibited LPS-induced BV2 cell synapse shortening, iNOS, and COX-2 protein expression more significantly than F16 and had stronger anti-inflammatory effects. On the other hand, in the oxidative damage mode induced by H2O2-induced BV2 cells, F16 activated HO-1 protein more than F17, which had a stronger antioxidant effect. |
