白花败酱草总黄酮提取工艺优化及抗腹泻活性研究
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摘要
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.
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.
引文
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[2]徐绳武,余安安,郑丽,等.白花败酱茶食两用产品的制作工艺[J].湖北农业科学, 2018,(21):91-92.
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[7]崔文燕,刘素香,宋晓凯,等.黄花败酱草和白花败酱草的化学成分与药理作用研究进展[J].药物评价研究,2016, 39(3):482-488.
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[10]王莉萍,李武,胡奕军,等.白花败酱总黄酮对小鼠抗氧化作用的影响[J].江西医药, 2015, 50(9):957-958, 967.
[11]黄素华,林标声,邱丰艳,等.白花败酱草总黄酮的提取及功能活性研究[J].龙岩学院学报, 2016, 34(5):117-119, 136.
[12]库咏峰,黄品鲜,陈萍,等.肉桂总黄酮提取工艺及其抗氧化性研究[J].应用化工, 2011, 40(9):1547-1552.
[13]崔文燕,刘素香,赵艳敏,等.响应曲面法优化黄花败酱总皂苷提取工艺及体外促进肠平滑肌收缩活性[J].中草药, 2016, 47(12):2078-2083.
[14]张建英,左爱学,刘羿晨.升麻萃取物对大鼠离体肠肌运动的影响研究[J].中国药业, 2015, 24(23):16-17.
[15]刘红艳,郭建生,王小娟,等.檀香不同部位提取物对豚鼠离体肠平滑肌功能的影响[J].西安交通大学学报(医学版), 2012, 33(2):241-244.
[16]朱明辉,邹佩,王桂华,等.吊篮式提取与传统提取方式所得健胃消食片浸膏粉对老鼠胃排空及小肠推进的影响研究[J].机电信息, 2017(32):41-45.
[17]王婷婷,钟凌云,徐婷.不同姜汁炮制黄连对小鼠止泻作用及胃肠动力的影响[J].时珍国医国药, 2017, 28(8):1876-1878.
[18]李杰,赵雨,普玲玲,等.生姜黄酮提取工艺优化研究[J].食品研究与开发, 2018, 39(22):59-65.
[19]江苏新医学院.中药大辞典[M].上海:上海人民出版社, 1975:1340-1341.
[20]刘竹凤,刘竹华.白头翁止泻汤肛门滴入治疗慢性腹泻126例[J].陕西中医, 2005(12):1331-1332.
[21]牛明了,甄欢欢.败酱草为主治疗溃疡性结肠炎38例临床观察[J].新中医, 2012, 44(12):55-56.
[2]徐绳武,余安安,郑丽,等.白花败酱茶食两用产品的制作工艺[J].湖北农业科学, 2018,(21):91-92.
[3]曾祥玲,王丹,邹晶晶,等.不同工艺条件白花败酱茶的品质比较分析[J].现代食品科技, 2019, 35(1):157-164, 232.
[4]李娜.白花败酱草化学成份、抗炎作用以及总皂苷提取纯化工艺研究[D].成都:成都中医药大学, 2006.
[5]万德安,杜成.白花败酱草治疗流行性腮腺炎[J].上海中医药杂志, 1985, 11.
[6]万新,石晋丽,刘勇,等.败酱属植物化学成分与药理作用[J].现代药物临床, 2006, 21(2):53-59.
[7]崔文燕,刘素香,宋晓凯,等.黄花败酱草和白花败酱草的化学成分与药理作用研究进展[J].药物评价研究,2016, 39(3):482-488.
[8]阎新佳,郑威,温静,等.白花败酱草的化学成分研究[J].中草药, 2017, 48(2):247-251.
[9]徐洁昕,周方钦.白花败酱中总黄酮的提取研究[J].广州食品工业科技, 2004(4):42-43.
[10]王莉萍,李武,胡奕军,等.白花败酱总黄酮对小鼠抗氧化作用的影响[J].江西医药, 2015, 50(9):957-958, 967.
[11]黄素华,林标声,邱丰艳,等.白花败酱草总黄酮的提取及功能活性研究[J].龙岩学院学报, 2016, 34(5):117-119, 136.
[12]库咏峰,黄品鲜,陈萍,等.肉桂总黄酮提取工艺及其抗氧化性研究[J].应用化工, 2011, 40(9):1547-1552.
[13]崔文燕,刘素香,赵艳敏,等.响应曲面法优化黄花败酱总皂苷提取工艺及体外促进肠平滑肌收缩活性[J].中草药, 2016, 47(12):2078-2083.
[14]张建英,左爱学,刘羿晨.升麻萃取物对大鼠离体肠肌运动的影响研究[J].中国药业, 2015, 24(23):16-17.
[15]刘红艳,郭建生,王小娟,等.檀香不同部位提取物对豚鼠离体肠平滑肌功能的影响[J].西安交通大学学报(医学版), 2012, 33(2):241-244.
[16]朱明辉,邹佩,王桂华,等.吊篮式提取与传统提取方式所得健胃消食片浸膏粉对老鼠胃排空及小肠推进的影响研究[J].机电信息, 2017(32):41-45.
[17]王婷婷,钟凌云,徐婷.不同姜汁炮制黄连对小鼠止泻作用及胃肠动力的影响[J].时珍国医国药, 2017, 28(8):1876-1878.
[18]李杰,赵雨,普玲玲,等.生姜黄酮提取工艺优化研究[J].食品研究与开发, 2018, 39(22):59-65.
[19]江苏新医学院.中药大辞典[M].上海:上海人民出版社, 1975:1340-1341.
[20]刘竹凤,刘竹华.白头翁止泻汤肛门滴入治疗慢性腹泻126例[J].陕西中医, 2005(12):1331-1332.
[21]牛明了,甄欢欢.败酱草为主治疗溃疡性结肠炎38例临床观察[J].新中医, 2012, 44(12):55-56.