College of Landscape Architecture and Art, Northwest A & F University, Yangling 712100, China.
Shanghai United International School, Gubei Secondary Campus, Shanghai 201103, China.
Molecules. 2022 Jul 4;27(13):4299. doi: 10.3390/molecules27134299.
Red 'Royalty' fruits are rich in anthocyanins. This study aimed to obtain the optimal parameters for the extraction and separation of anthocyanins from 'Royalty' fruits and to evaluate the inhibitory effect of the enriched anthocyanin fraction on gastric cancer cells. Ultrasonic-assisted extraction was used for the extraction of the anthocyanins of Malus 'Royalty' fruit, and the extraction results showed that the optimum parameters were an extraction temperature of 20 °C, a solid-liquid ratio of 1:6 (g/mL), ethanol and formic acid contents of 70% and 0.4%, respectively, an extraction time of 40 min, and an ultrasonic power of 300 W. The optimum extraction parameters to achieve the highest anthocyanin yield by a single-factor experiment coupled with response surface methodology were identified. The separation results showed that the AB-8 macroporous resin was a better purifying material, with 60% ethanol as an adsorbent, and the adsorption-desorption equilibrium times were 6 h and 1 h, respectively. Cyanidin-3-galactoside was the main body composition separation of anthocyanins by a high-performance liquid chromatography-diode array detector. The antitumor activity results showed that the anthocyanins of 'Royalty' fruits have a significant inhibitory effect on the gastric cancer cell line BGC-803. The in vitro cell viability test of CCK-8 showed that the inhibitory effect on tumor cells was more significant with the increased anthocyanin concentration, with a half maximal inhibitory concentration (IC) value of 105.5 μg/mL. The cell morphology was observed by an inverted microscope, and it was found that the backbone of BGC-803 treated with a high concentration of anthocyanins was disintegrated and the nucleoplasm was concentrated. The mechanism of apoptosis was analyzed by Western blotting, and the results showed that with increasing anthocyanin concentration in the medium, the expression levels of the proapoptotic proteins Bax and Bak increased, and the expression levels of the antiapoptotic proteins Bcl-2 and Bcl-xL decreased, which coordinated the regulation of cell apoptosis. This research suggests that the enriched anthocyanin fraction from 'Royalty' fruits have potential antitumor and adjuvant therapeutic effects on gastric cancer.
红‘贵族’水果富含花青素。本研究旨在获得从‘贵族’水果中提取和分离花青素的最佳参数，并评估富化花青素级分对胃癌细胞的抑制作用。采用超声波辅助提取法提取苹果‘贵族’果实中的花青素，提取结果表明，最佳参数为提取温度 20°C，固液比 1:6（g/mL），乙醇和甲酸含量分别为 70%和 0.4%，提取时间 40 min，超声功率 300 W。通过单因素实验与响应面法相结合确定了获得最高花青素产量的最佳提取参数。分离结果表明，AB-8 大孔树脂是一种较好的纯化材料，以 60%乙醇为吸附剂，吸附-解吸平衡时间分别为 6 h 和 1 h。高效液相色谱-二极管阵列检测器分离的花青素主要成分为矢车菊-3-半乳糖苷。体外细胞活力试验 CCK-8 表明，‘贵族’果实中的花青素对胃癌细胞系 BGC-803 具有显著的抑制作用。随着花青素浓度的增加，对肿瘤细胞的抑制作用更加明显，半数最大抑制浓度（IC）值为 105.5 μg/mL。倒置显微镜观察细胞形态，发现高浓度花青素处理的 BGC-803 细胞骨架崩解，核质浓缩。通过 Western blot 分析细胞凋亡的机制，结果表明，随着培养基中花青素浓度的增加，促凋亡蛋白 Bax 和 Bak 的表达水平增加，而抗凋亡蛋白 Bcl-2 和 Bcl-xL 的表达水平降低，协调细胞凋亡的调节。本研究表明，‘贵族’果实中富化的花青素级分对胃癌具有潜在的抗肿瘤和辅助治疗作用。
