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This study aimed to optimize the ultrasound-assisted macroporous resin purification process for Abelmoschus manihot (L.) leaf flavonoids (AMLF) and investigate their regulatory effect on intestinal microbiota in mice. Optimal adsorption rates 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 84.65%. 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.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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