超声协同天然低共熔溶剂法提取明日叶查尔酮工艺优化及其抗衰老作用研究

目的 优化超声协同天然低共熔溶剂法提取明日叶查尔酮的工艺, 并研究其抗衰老作用。方法 以明日叶查尔酮得率为指标, 采用单因素和响应面实验, 优化超声协同天然低共熔溶剂法的提取工艺条件, 并设计了水提法、醇提法、微波辅助法、超声辅助法与超声协同天然低共熔溶剂法的对比实验。同时将小鼠随机分为正常对照组、衰老模型组、维生素C (vitamin C, VC)阳性对照组、明日叶查尔酮低、中、高剂量组。除正常对照组外, 其余5组注射D-半乳糖构建衰老小鼠模型。测定小鼠血清中超氧化物歧化酶(superoxide dismutase, SOD)、总抗氧化能力(total antioxidant capacity, T-AOC)和丙二醛(malondialdehyde, MDA)含量、脏器指数和小鼠皮肤组织中的SOD、过氧化氢酶(catalase, CAT)、三磷酸腺苷酶(adenosine triphosphatase, ATPase)活性和脂褐素(lipofuscin, LPF)、羟脯氨酸(hydroxyproline, HYP)、Ⅰ型胶原蛋白含量。结果 最佳提取工艺条件为: 液固比30:1 (mL/g)、超声功率400 W、超声时间35 min, 此条件下明日叶查尔酮得率为0.94%, 此法的提取效果优于其他4种单一的提取方法。明日叶查尔酮对D-半乳糖致衰老小鼠的抗衰老实验结果表明: 明日叶查尔酮能有效增强小鼠血清中SOD、T-AOC活性, 降低MDA含量, 提高各脏器指数, 增强小鼠皮肤组织中SOD、CAT和ATPase活性, 及HYP和Ⅰ型胶原蛋白含量, 减少LPF的生成。结论 该研究获得了明日叶查尔酮的最佳提取工艺, 得到的明日叶查尔酮对D-半乳糖致衰小鼠具有较强的抗衰老作用。

Optimization of extraction process of Angelica keiskei chalcones by ultrasonic assisted natural deep eutectic solvents and its anti-aging effect

Objective To optimize the extraction process of Angelica keiskei chalcones by ultrasonic assisted natural deep eutectic solvents method and study its anti-aging effect. Methods Using the yield of Angelica keiskei chalcones as an indicator, single factor and response surface experiments were used to optimize the extraction process conditions of the ultrasonic assisted natural deep eutectic solvents method. The comparative experiments were designed for water extraction, alcohol extraction, microwave assisted extraction, ultrasound assisted extraction, and ultrasonic assisted natural deep eutectic solvents extraction. At the same time, the mice were randomly divided into normal control group, aging model group, vitamin C (VC) positive control group, and low, medium, and high dose of Angelica keiskei chalcones groups. In addition to the normal control group, the other 5 groups were injected D-galactose to build the aging mouse model. The superoxide dismutase (SOD), total antioxidant capacity (T-AOC) activity andmalondialdehyde (MDA) content in mouse serum, organ index, and SOD, catalase (CAT), adenosine triphosphatase (ATPase) activity and lipofuscin (LPF), hydroxyproline (HYP), and type I collagen content in mouse skin tissue were measured. Results The optimum extraction conditions were: Liquid-solid ratio of 30:1 (mL/g), ultrasound power of 400 W, and ultrasound time of 35 minutes. Under these conditions, the yield of Angelica keiskei chalcones was 0.94%, and the extraction effect of this method was superior to the other 4 single extraction methods. The anti-aging test results of Angelica keiskei chalcones on aging mice induced by D-galactose showed that Angelica keiskei chalcones could effectively enhance the activities of SOD and T-AOC in the serum of mice, reduce the content of MDA, increase the index of various organs, increase the activities of SOD, CAT and enhance the activity ATPase, increase the content of HYP and type I collagen in the skin tissue of mice, and reduce the production of LPF. Conclusion The study obtains the best extraction process of Angelica keiskei chalcones, and the obtained Angelica keiskei chalcones has a strong anti-aging effect on aging mice induced by D-galactose.