紫锥菊根中菊苣酸提取工艺及对氧化应激肠道细胞的保护作用研究

doi:10.16431/j.cnki.1671-7236.2022.07.007

摘要/Abstract

摘要： 【目的】优化紫锥菊根中菊苣酸提取工艺，并测定菊苣酸在肠道细胞中的抗氧化效果。【方法】试验选用有机溶剂乙醇作为紫锥菊根粉末（80目）提取溶剂，对提取溶剂浓度、提取温度、提取时间、提取次数以及原料与溶剂的比例等因素进行分析，得到每个因素下菊苣酸得率和该因素的对应曲线，再在单因素基础上，选择溶剂浓度（30%、40%、50%）、提取温度（60、70、80 ℃）、提取时间（1、2、3 h）和料液比（1∶12、1∶15、1∶18）4个因素3个水平设计正交试验。用不同浓度H₂O₂（100、200、400、600、800、1 000 μmol/L）处理人结肠腺癌细胞（Caco-2）与猪小肠上皮细胞（IPEC-J2）建立氧化应激模型。分别使用不同浓度菊苣酸（0、100、200、400、600、800、1 000 μmol/L）处理氧化应激前后的细胞24 h，应用MTT法，分别测定菊苣酸对两种肠道细胞氧化应激的保护和缓解效果。【结果】菊苣酸最佳提取条件为：50%浓度的乙醇在70 ℃下采用1∶18的料液比，超声辅助提取2 h，得率为1.89 mg/g。细胞抗氧化试验结果显示，菊苣酸对Caco-2和IPEC-J2两种肠道细胞氧化应激均具有保护和缓解效果，对Caco-2细胞的保护作用和缓解作用的半数抑制浓度（IC₅₀）值分别为172.6和207.5 μmol/L，对IPEC-J2细胞的保护作用和缓解作用的IC₅₀值分别为133.1和196.5 μmol/L。【结论】试验得到了紫锥菊根粉中菊苣酸的最佳提取方案，为工业化高效提取菊苣酸提供了参考；同时菊苣酸对氧化应激的肠道细胞具有很好的保护作用，经计算其保护作用的IC₅₀值较缓解效果更小，进一步证明了菊苣酸具有成为肠道抗氧化剂的潜力。

关键词: 紫锥菊; 菊苣酸; 提取; 肠道细胞; 抗氧化作用

Abstract: 【Objective】 The extraction process of cichoric acid from Echinacea root was optimized,and the antioxidant effect of cichoric acid in intestinal cells was determined.【Method】 In the experiment,the economical organic solvent ethanol was selected as the extraction solvent of Echinacea root powder (80 mesh),and several factors such as extraction solvent concentration,extraction temperature,extraction time,extraction times,and the ratio of raw materials to solvent were set for analysis,and the results of each factor were obtained.The corresponding curve of acid yield and this factor,and then on the basis of single factor,select solvent concentration (30%,40%,50%),extraction temperature (60,70,80 ℃),extraction time (1,2,3 h),solid-liquid ratio (1∶12,1∶15,1∶18),four factors and three levels of orthogonal experiments were designed.With different concentrations of H₂O₂ (100,200,400,600,800,1 000 μmol/L) to treat human colon adenocarcinoma cells (Caco-2) and porcine small intestinal epithelial cells (IPEC-J2).The cells before and after oxidative stress were treated with chicory acid (0,100,200,400,600,800,1 000 μmol/L) for 24 h respectively.The protective and alleviating effects of chicory acid on oxidative stress of two intestinal cells were measured by MTT method.【Result】 The results of extraction process showed the optimal extraction conditions of chicoric acid were 50% ethanol at 70 ℃ with a solid-liquid ratio of 1∶18,ultrasonic-assisted extraction for 2 h.Finally,the yield of this process was 1.89 mg/g.The results of cellular antioxidant assay showed that chicoric acid had preventive and alleviating effects on both Caco-2 and IPEC-J2 cell lines which damaged by oxidative stress.The IC₅₀ values of its protective and alleviating effects were 172.6 and 207.5 μmol/L for Caco-2,133.1 and 196.5 μmol/L for IPEC-J2,respectively.【Conclusion】 The best extraction process of chicoric acid from Echinacea root powder was obtained,which provided a reference for industrial and efficient extraction of chicoric acid,and the cell antioxidant experiment showed the intestinal antioxidant ability of chicoric acid.The calculated IC₅₀value of its protective effect was smaller than that of the mitigation effect.It was further proved that chicoric acid had the potential as a future intestinal antioxidant drug.

Key words: Echinacea; chicoric acid; extraction; intestinal cells; antioxidant effect

中图分类号:

S859.7

李一鹏, 张虎, 任娟, 任欣, 刘聚祥, 王庚南. 紫锥菊根中菊苣酸提取工艺及对氧化应激肠道细胞的保护作用研究[J]. 中国畜牧兽医, 2022, 49(7): 2497-2505.

LI Yipeng, ZHANG Hu, REN Juan, REN Xin, LIU Juxiang, WANG Gengnan. Study on the Extraction Process of Chicoric Acid from Echinacea Root and Its Protective Effects on Oxidative Stress Damaged Intestinal Cells[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2497-2505.

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