基于药效筛选和正交试验优化五子衍宗加减方的提取工艺
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摘要
目的通过长期毒性与药效实验筛选五子衍宗加减方(WYR)的提取工艺,并以正交试验进行优化,确定WYR的最佳提取工艺。方法对不同工艺WYR样品进行长期毒性实验,并以肾阳虚大鼠阴茎勃起潜伏期和性激素水平为药效指标,综合长期毒性与药效实验的结果,筛选出最佳工艺方案;以多个指标成分含量和出膏率为指标,采用正交试验筛选不同提取工艺的最优水平,并进行验证。结果长期毒性实验结果显示所有小鼠均健康存活,未见明显毒性反应。药效实验结果表明,巴戟天、杜仲、桑寄生、续断、炙甘草以70%乙醇回流提取,其余药材用70%乙醇作为溶剂进行渗漉提取为WYR最佳提取方式;得到的最优提取工艺为巴戟天、杜仲、桑寄生、续断、炙甘草以10倍量70%乙醇回流提取3次,每次2.0 h;其余药材以10倍量70%乙醇1.5 mL/min渗漉提取,渗漉前药材先浸泡16 h。结论 WYR能明显改善肾阳虚大鼠阴茎勃起潜伏期和性激素水平,优选的工艺条件合理可行,为WYR的后续开发提供依据。
Objective To screen the extraction process of Wuzi Yanzong plus and minus Recipe(WYR) by pharmacodynamics and long-term toxicity test, and optimize it by orthogonal experiment to determine the best extraction process of WYR. Methods Long-term toxicity experiments were carried out on different processed WYR samples. The penile erection latency and sex hormone levels in rats with kidney-yang deficiency were used as pharmacodynamic indicators. The results of long-term toxicity and efficacy experiments were combined to screen out the optimal process plan. Using the content of ingredients and the rate of ointment as indicators, the orthogonal test was used to screen the optimal level of different extraction processes and verified. Results Long-term toxicity test results showed that all mice survived healthily and no obvious toxicity was observed. The results of pharmacodynamic experiments showed that the extracts of Morindae Officinalis Radix, Eucommiae Cortex, Taxilli Herba, Dipsaci Radix, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were refluxed with 70% ethanol, and the remaining medicinal materials were extracted with 70% ethanol as the solvent to obtain the best extraction method of WYR. The suitable extraction process was as follows:Morindae Officinalis Radix, Eucommiae Cortex, Taxilli Herba, Dipsaci Radix, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were extracted three times with 10 times 70% ethanol for 2.0 h each time; The remaining medicinal materials were percolated with 10 times 70% ethanol at 1.5 mL/min. The medicinal herbs were firstly soaked for 16 h before percolation. Conclusion WYR can significantly improve the penile erection latency and sex hormone levels in rats with kidney yang deficiency. The optimal process conditions are reasonable and feasible, which provides a basis for the follow-up development of WYR.
Objective To screen the extraction process of Wuzi Yanzong plus and minus Recipe(WYR) by pharmacodynamics and long-term toxicity test, and optimize it by orthogonal experiment to determine the best extraction process of WYR. Methods Long-term toxicity experiments were carried out on different processed WYR samples. The penile erection latency and sex hormone levels in rats with kidney-yang deficiency were used as pharmacodynamic indicators. The results of long-term toxicity and efficacy experiments were combined to screen out the optimal process plan. Using the content of ingredients and the rate of ointment as indicators, the orthogonal test was used to screen the optimal level of different extraction processes and verified. Results Long-term toxicity test results showed that all mice survived healthily and no obvious toxicity was observed. The results of pharmacodynamic experiments showed that the extracts of Morindae Officinalis Radix, Eucommiae Cortex, Taxilli Herba, Dipsaci Radix, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were refluxed with 70% ethanol, and the remaining medicinal materials were extracted with 70% ethanol as the solvent to obtain the best extraction method of WYR. The suitable extraction process was as follows:Morindae Officinalis Radix, Eucommiae Cortex, Taxilli Herba, Dipsaci Radix, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were extracted three times with 10 times 70% ethanol for 2.0 h each time; The remaining medicinal materials were percolated with 10 times 70% ethanol at 1.5 mL/min. The medicinal herbs were firstly soaked for 16 h before percolation. Conclusion WYR can significantly improve the penile erection latency and sex hormone levels in rats with kidney yang deficiency. The optimal process conditions are reasonable and feasible, which provides a basis for the follow-up development of WYR.
引文
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[2]戎志斌,郭文鹏,赵志英,等.五子衍宗方合逍遥散加减对男子弱精症的临床疗效[J].微循环学杂志,2015,25(2):37-40.
[3]李媛,杨智.马居里运用五子衍宗方加减治疗男科疾病[J].河南中医,2013,33(3):401-402.
[4]徐玉玲,白婷钰,付春梅,等.三七配伍丹参对丹参中成分提取溶出的影响[J].中草药,2016,47(24):4357-4363.
[5]殷明阳,刘毅,那溪元,等.正交试验优化止咳平喘颗粒处方药材的乙醇提取工艺[J].现代药物与临床,2015,30(10):1208-1211.
[6]张周菊,张悦,彭翠香,等.药效学结合正交试验优选蛇归止痒方的提取工艺[J].中国药房,2018,29(19):2662-2666.
[7]余姣,陈娇,周文杰,等.基于抗炎解热药效的小儿温热口服液中心复合设计-效应面法提取工艺优化研究[J].中草药,2016,47(20):3632-3638.
[8]Li Y,Zhu C P,Zhai X C,et al.Optimization of enzyme assisted extraction of polysaccharides from pomegranate peel by response surface methodology and their antioxidant potential[J].Chin Herb Med,2018,10(4):416-423.
[9]崔婷婷,黄晴雯,李心雨,等.2次旋转正交设计优化锁阳总糖提取工艺[J].中草药,2016,47(22):4003-4008.
[10]石浩源,王春雷,赵艳敏,等.正交试验结合体外抑制肠平滑肌收缩活性研究优化固公果根总三萜提取工艺[J].药物评价研究,2017,40(11):1581-1586.
[11]陈剑平,陈秋谷,胡兆流,等.基于药效筛选和正交试验优化健脾益肾丸的制备工艺[J].中草药,2018,49(24):5817-5822.
[12]芦彦兆.基于化学与药效学结合的藏药湿生扁蕾的提取方法及工艺的研究[D].兰州:甘肃中医药大学,2018.
[13]庞燕梅,周福军,胡跃强,等.基于药效筛选和正交试验优化温肺降浊方的提取工艺[J].中草药,2018,49(10):2420-2425.
[14]徐继建,丁志勇,昊依娜,等.五子衍宗丸长期毒性实验研究[J].湖南中医药导报,2001,7(6):333-335.
[15]葛争艳,金龙,刘建勋.五子衍宗丸补肾壮阳作用的实验研究[J].中国实验方剂学杂志,2010,16(7):173-176.
[16]徐叔云,卞如濂,陈修.药理实验方法学[M].第3版.北京:人民卫生出版社,1982.
[17]中国药典[S].一部.2015.