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To optimize the water extraction process of modified Tabusen-2 by response surface methodology (Box-Behnken) and investigate its in vitro efficacy.
Taking the mass fractions of caffeic acid and geniposide as the comprehensive score, response surface methodology was used to optimize the soaking time, material-to-liquid ratio, extraction time and extraction times. The RAW 264.7 cells induced by nuclear factor-κB receptor activator ligand (RANKL) were co-cultured with the optimal extracts for 5 d, and then treated with tartrate-resistant acid phosphatase (TRAP). The TRAP-positive cells were stained and observed under a microscope to evaluate the in vitro efficacy of the extract.
The extraction time, soaking time, and material-to-liquid ratio were selected as the inspection factors, and the comprehensive score of caffeic acid and geniposide content was used as the evaluation index. After optimization by response surface method, the optimal extraction condition was 540 min; the liquid ratio was 1∶12 (g·mL-1); the extraction time was 44 min, and the extraction times were two times. The extract obtained under this condition had a significant inhibitory effect on RAW264.7 cells induced by RANKL, could reduce TRAP-positive cells, and inhibited multinucleation.
The water extraction process of modified Tabusen-2 optimized by response surface analysis method is reproducible, stable and feasible, and has good in vitro pharmacodynamic activity, laying a reliable foundation for future in vivo pharmacodynamic experimental research.
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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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