摘要
目的:筛选与动物药功能主治相关活性组分,探讨其传统热炮制加工机理,验证肽键热振荡理论。方法:以4类动物药为代表,包括虫类、胶类、角类、贝壳类,对动物药连续用石油醚、乙酸乙酯、甲醇、水超声提取,得到4个不同极性溶媒提取物,对其进行系统活性对比研究。进一步将动物药水提液经超滤技术分为>10kDa、3~10kDa、1~3kDa、<1kDa组分。以相关活性指标,筛选活性组分;对炮制前后动物药<1kDa组分的活性及寡肽收率进行对比研究;以动物药蛋白>10kDa部分为底物进行酶解,以相关活性指标对天然和酶解<1kDa组分进行对比研究。结果:动物药各极性溶媒提取物活性最强的为水提取物,水提取物各组分中<1kDa或1~3kDa的小肽段活性明显强于蛋白、多肽部位,因此确定动物药<1kDa或1~3kDa的小肽段为活性组分;炮制品的寡肽含量高于生品;酶解<1kDa组分活性略低于天然<1kDa组分,但酶解寡肽的寡肽收率远高于天然寡肽。结论:确定了<1kDa或1~3kDa的小肽段组分为动物药的药效物质基础;动物药经炮制之后,由于肽键的断裂,生成了更多的小分子寡肽类成分,进而可以发挥更强活性;进一步阐明了动物药热炮制机理,验证了课题组提出的肽键热振荡理论;此外,由蛋白酶解得到更多的活性寡肽,解决了天然寡肽收率较低难于产业化的问题,为动物药小肽段类物质开发应用提供了可靠的依据。
Objective: To screen the active components related to the main functions of animal drugs,explore the mechanism of traditional thermal processing,and verify the theory of thermal oscillation of peptide bonds. Methods: Four kinds of animal medicines,including insects,colloids,horns and testacean,were extracted by ultrasound with petroleum ether ,ethyl acetate,methanol and water.Four different polar solvent extracts were obtained and their activities were compared.Ultrafiltration technology was used to divide the aqueous extracts of animal medicines into four components:more than 10 kDa,3-10 kDa,1-3 kDa and less than 1 kDa,and the active components were screened by the related activity indexes.The activity and oligopeptide yield of less than 1 kDa component of animal medicine before and after processing were compared.The enzymatic hydrolysis was carried out on the part of animal drug greater than 10 kDa,and the natural and enzymatic components less than 1 kDa were compared with the related activity indexes. Result: The most active component of the polar solvent extracts of animal medicine was aqueous extract.The activity of small molecular weight peptides less than 1 kDa or 1-3 kDa in each component of aqueous extract was obviously stronger than that of protein and polypeptide parts.Therefore,the small molecular weight peptides of animal medicine less than 1 kDa or 1-3 kDa were determined as active components,and the oligopeptide content of processed products was higher than that of raw products.The activity of oligopeptides with enzymatic hydrolysis less than 1 kDa were slightly lower than those with natural hydrolysis less than 1 kDa,but the yield of oligopeptides with enzymatic hydrolysis was much higher than that of natural oligopeptides. Conclusion: The small molecular weight peptides of 1 kDa or 1-3 kDa were identified as the basis of pharmacodynamic substances of animal drugs,and after processing more small molecular oligopeptides were produced due to the breakage of peptide bonds,which could play a more active role.The mechanism of thermal processing of animal drugs was further clarified,and the theory of thermal oscillation of peptide bonds proposed by the research group was verified.In addition,more active oligopeptides were obtained by proteolysis,which may solve the problem that the yield of natural oligopeptides is low and difficult to industrialize,and it provides a reliable basis for the development and application of small molecular peptides in animal medicine.
作者
李晶峰
边学峰
孙佳明
林喆
张辉
LI Jing-feng;BIAN Xue-feng;SUN Jia-ming;LIN Zhe;ZHANG Hui(Changchun University of Traditional Chinese Medicine,Changchun 130117,China)
出处
《中国现代中药》
CAS
2019年第9期1147-1156,共10页
Modern Chinese Medicine
基金
公益性行业科研专项(201507002)
国家自然基金面上项目(81373936)
关键词
肽键热振荡理论
酶工程技术
动物药
肽类化合物
thermal oscillation theory of peptide bond
enzyme engineering technology
animal medicine
peptide compounds
