超声辅助大孔树脂纯化菜芙蓉叶黄酮及其对肠道菌群的影响

蔡梦权; 杨胜男; 熊小英; 郑明洋; 赵斗; 仇燕

doi:10.13386/j.issn1002-0306.2025070250

超声辅助大孔树脂纯化菜芙蓉叶黄酮及其对肠道菌群的影响

Ultrasound-assisted Macroporous Resin Purification of Flavonoids from Ambelmoschus manihot (L.) Leaves and Their Effect on Mice Gut Microbiota

摘要

摘要: 本研究旨在优化超声辅助大孔树脂技术纯化菜芙蓉叶黄酮（Ambelmoschus manihot (L.) leaves flavonoid，AMLF）工艺，并探究AMLF对小鼠肠道菌群与代谢产物的影响。通过超声功率、温度、时间和上样浓度等因素确定最佳吸附率，并以超声功率、温度、时间和乙醇体积分数确定最佳解吸率，结合吸附率和解吸率双指标，确定最佳纯化工艺。连续灌胃正常小鼠不同浓度AMLF（80、160、320 mg/kg）35 d，测定小肠总抗氧化能力（Total Antioxidant Capacity，T-AOC），过氧化氢酶（Catalase，CAT），丙二醛（Malondialdehyde，MDA），超氧化物歧化酶（Superoxide dismutase，SOD）和分泌型免疫球蛋白A（Secretory immunoglobulin A，sIgA）含量，测定血清组织白细胞介素1β（Interleukin-1β，IL-1β）、白细胞介素2（Interleukin-2，IL-2）、免疫球蛋白G（Immunoglobulin G，IgG）的含量，分析肠道菌群组成以及粪便短链脂肪酸（Short-chain fatty acids，SCFAs）含量。结果表明，选用HPD500大孔树脂，在超声功率150 W，温度25 ℃，时间60 min，上样浓度1.3 mg/mL的吸附条件下，黄酮吸附率最高达51.76%；在超声功率180 W，温度45 ℃，时间30 min，乙醇体积分数70%的解吸条件下，解吸率达84.64%，在此工艺下AMLF纯度可达36.72%。与空白组相比，高剂量组（320 mg/kg）小鼠CAT含量提高了67.43%（P<0.05），而MDA含量则降低了50.41%（P<0.05）。AMLF能够提高小鼠肠道菌群多样性，上调有益菌Lactobacillus和Prevotellaceae_UCG_001相对丰度，促进SCFAs的产生。综上，超声辅助大孔树脂技术绿色低耗、易于放大，可显著增加AMLF的纯度，且AMLF能够调节机体免疫能力，提高抗氧化能力，改变肠道菌群结构与代谢产物种类，促进肠道健康，为菜芙蓉叶资源的开发及其在食品领域的应用提供理论基础。

Abstract: This study aimed to optimize the ultrasound-assisted macroporous resin purification process for Ambelmoschus manihot (L.) leaf flavonoids (AMLF) and investigate their regulatory effect on intestinal microbiota in mice. Optimal adsorption conditions were determined by evaluating ultrasound power, temperature, time, and sample loading concentration, while desorption efficiency was optimized via ultrasound power, temperature, time, and ethanol volume fraction. The final purification protocol was established by integrating both adsorption and desorption performance. The normal mice were gavaged with AMLF at doses of 80, 160, or 320 mg/kg for 35 days. Biomarkers including total antioxidant capacity (T-AOC), catalase (CAT), malondialdehyde (MDA), superoxide dismutase (SOD), and secretory immunoglobulin A (sIgA) in intestinal tissues, as well as serum interleukin-1β (IL-1β), interleukin-2 (IL-2), and immunoglobulin G (IgG) levels, were measured. Intestinal microbiota composition and fecal short-chain fatty acids (SCFAs) were analyzed respectively. The results indicated that HPD500 macroporous resin under adsorption conditions (150 W ultrasound power, 25 ℃, 60 min, 1.3 mg/mL loading concentration) achieved the highest adsorption rate (51.76%). Optimal desorption was attained at 180 W ultrasound power, 45 ℃, 30 min, and 70% ethanol, yielding a desorption rate of 84.64%. Under this protocol, AMLF purity increased to 36.72%. Compared with the control group, the intestinal CAT activity in mice of the high-dose group (320 mg/kg) was significantly elevated by 67.43% (P<0.05), whereas the MDA content was markedly reduced by 50.41% (P<0.05). AMLF treatment enhanced intestinal microbiota diversity, upregulated the relative abundance of beneficial bacteria (e.g., Lactobacillus and Prevotellaceae_UCG_001), and promoted SCFA production. In summary, the ultrasound-assisted macroporous resin technology, characterized by its greenness, low energy consumption, and scalability, effectively enhances the purity of AMLF. Moreover, AMLF modulates immune function, enhances antioxidant capacity, alters the structure of intestinal microbiota and the composition of metabolites, and promotes intestinal health. These findings collectively lay a theoretical foundation for the development of A. manihot (L.) leaf resources and their application in the food industry.