白花败酱草总黄酮提取工艺优化及抗腹泻活性研究

Study on extracting technology optimization and antidiarrheal activity of total flavonoids from Patrinia villosa

目的对白花败酱草中总黄酮的提取工艺和抗腹泻活性进行初步研究。方法以芦丁为对照品,以单位质量生药中总黄酮含量为指标,采用硝酸铝显色比色-紫外分光光度法,通过单因素考察和响应面设计优化有机溶剂法提取白花败酱草茎总黄酮的工艺,分别考察乙醇浓度、料液比、提取时间和提取温度对白花败酱草总黄酮(PTF)提取效果的影响。通过小鼠小肠酚红推进实验和大鼠离体肠平滑肌实验评估白花败酱草总黄酮在体内、外对腹泻模型中肠动力的影响。结果经优化,在溶剂为60%乙醇、料液比1∶8 g/mL、85℃提取2.5 h的条件下,白花败酱草茎的总黄酮含量可达25 mg/g以上。白花败酱草总黄酮可以显著降低乙酰胆碱诱发的收缩亢进的大鼠离体肠平滑肌收缩张力,且对不同肠段的作用强度顺序为:空肠>十二指肠>回肠>结肠。与模型组比较,白花败酱草总黄酮能够明显抑制酚红在蓖麻油致腹泻小鼠小肠中的推进进程(P<0.05、0.01)。结论有机溶剂法提取白花败酱草茎中总黄酮的工艺通过响应曲面分析得到优化,白花败酱草总黄酮可以抑制肠蠕动,具有潜在的抗腹泻活性。

Objective To optimize the extracting conditions of total flavonoids from Patrinia villosa stem and to preliminarily investigate the antidiarrheal activity of total flavonoids from P. villosa. Methods UV spectrophotometry was used to determine the content of total flavonoids by taking rutin as the standard substance, aluminum nitrate as the chromogenic agent and taking the content of total flavonoids in per gram of crude drug as the index. The extracting technologies were evaluated through signal factor study and response-surface-design experiments. The in vitro and in vivo effects of total flavonoids of P. villosa on intestinal motility in the diarrheal model were evaluated by isolated intestine test and testing the propelling of phenol red in small intestine of mice.Results The optimum extracting conditions of total flavonoids from P. villosa stem includes the extracting solvent of 60% ethanol,ratio of material to liquid at 1:8 g/mL, extracting time of 2.5 h and the temperature at 85 °C, under which condition the content of total flavonoids from P. villosa stem could reach over 25 mg/g. The total flavonoids of P. villosa can significantly reduce the contractile tension of isolated acetylcholine-induced hyperactive rat intestinal smooth muscles, and the order of effect on different intestinal segments was: jejunum>duodenum>ileum>colon. The total flavonoids of P. villosa significantly inhibit the progress of phenol red in the small intestine of castor oil-loaded diarrhea mice(P < 0.05, 0.01). Conclusion The organic solvent-extracting conditions of total flavonoids from the stems of P. villosa were optimized by response surface analysis. The total flavonoids of P.villosa inhibit intestinal peristalsis and were verified to possess the potential antidiarrheal activity.