木奶果葉萃取物之抗氧化活性及對人類大腸直腸癌細胞
Caco-2 抑制活性之研究

木奶果(Baccaurea ramiflora)為大戟科之藥用植物，於中國少數民族傣族的醫藥學中可作為消炎止痛藥。經研究顯示，木奶果葉片甲醇萃取物具有抗氧化、降血糖及抗發炎等功效。由於甲醇具有較強之毒性，且會使人體受到傷害，因此本研究將木奶果葉片以 95℃熱水及不同濃度乙醇 (25%、50%、75%、95%) 進行萃取後，分析生物活性成分、抗氧化活性及對人類大腸直腸癌細胞 Caco-2 之抑制活性。結果顯示，總酚以 75%乙醇萃取物含量最高達 354.6±0.764 mg/g；總類黃酮以 95%乙醇萃取含量為最高達 60.3±1.967 mg/g。經 HPLC-UV/Vis 分析，熱水含有較多酚類化合物，分別為綠原酸(3.86±0.07 mg/g)、兒茶素 (0.65±0.04 mg/g)、咖啡酸(1.63±0.01 mg/g)及槲皮素(5.95±0.02 mg/g)。於抗氧化試驗中，DPPH、ABTS 及還原能力以 75%乙醇萃取最佳，其 IC50 值分別為 15.732±0.309 g/mL 、168.236±10.97 g/mL 與 209.89±16.49 g/mL。於細胞存活試驗，75%乙醇萃取物於 800 g/mL 濃度下作用 72 小時後，抑制癌細胞存活率達 80 %以上，並透過流式細胞儀分析細胞週期及細胞凋亡，結果顯示 75%乙醇萃取物隨著濃度及時間的增加，增加 SubG1 期之細胞比例及總凋亡百分比。利用西方墨點法進一步分析細胞凋亡途徑中凋亡蛋白含量，75%乙醇萃取物透過增加 Bax 蛋白及降低 pro-caspase3 蛋白使細胞以內部粒線體途徑走向凋亡。綜上所述，木奶果葉片含有較高活性物質及抗氧化能力，且於大腸直腸癌細胞存活試驗，75%乙醇萃取物有較佳之細胞毒殺效果，並以濃度及時間依賴性方式增加 SubG1 期及總凋亡百分比，因此木奶果葉片有助於未來作為保健食品之開發。

關鍵字

木奶果葉；抗氧化活性；大腸直腸癌細胞；細胞凋亡

並列摘要

Baccaurea ramiflora is a medicinal plant of the Euphorbiaceae family, which is used as an anti-inflammatory and analgesic in the medicine of the Chinese minority Dai ethnic minority. Previous studies indicate that Baccaurea ramiflora Lour leaves extracted with methanol showed antioxidant, hypoglycemic, and anti-inflammatory effects. Since methanol is highly toxic and can cause damage to humans, in this study, Baccaurea ramiflora Lour leaves were extracted with hot water at 95°C and different concentrations with ethanol (25%, 50%, 75%, 95%) to analyze the bioactive components, antioxidant activity and inhibitory activity against human colorectal cancer cells Caco-2. The results showed that 75% ethanol extract demonstrated the highest total phenol content with 354.6±0.764 mg/g, while 95% ethanol extract exhibited the greatest flavonoid content with 60.3±1.967 mg/g. Following HPLC-UV/Vis analysis, The hot water contained more phenolic compounds, chlorogenic acid (3.86±0.07 mg/g), catechin (0.65±0.04 mg/g), caffeic acid (1.63±0.01 mg/g) and quercetin (5.95±0.02 mg/g). For the antioxidant assay, DPPH, ABTS, and reducing power showed the strongest IC50 values of 15.732±0.309 g/mL, 168.236±10.97 g/mL, and 209.89±16.49 g/mL for 75% ethanol extract. In cell experiments, 75% ethanol extract was concentrated at 800 g/mL for 72 hours, inhibiting the cancer cell viability up to 80%. The cell cycle and apoptosis were analyzed by flow cytometry, and the results showed that 75% ethanol extract increased the subG1 phase and total apoptosis percentages with the increase in concentration and time. Using western blots to further analyze apoptotic proteins in the apoptotic pathway, 75% ethanol extracts caused apoptosis by increasing Bax protein and decreasing pro-caspase3 protein in the internal pathway. Conclusively, Baccaurea ramiflora Lour leaves could be useful for the future development of healthy food.

並列關鍵字

Baccaurea ramiflora Lour leaves ； Antioxidant activity ； Colorectal carcinoma cells ； Cell apoptosis

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