基于造血活性小肽研究阿胶炮制原理  

Processing principles of Asini Corii Colla based on hematopoietic small peptides

作　　者：付英杰[1] 司子林 刘月 贾玉民 陈智[5] 陈小天 王建安[1] FU Ying-jie;SI Zi-lin;LIU Yue;JIA Yu-min;CHEN Zhi;CHEN Xiao-tian;WANG Jian-an(College of Pharmacy,Jining Medical University,Rizhao 276826,China;College of Pharmacy,China Pharmaceutical University,Nanjing 211198,China;College of Pharmacy,Jiangxi University of Traditional Chinese Medicine,Nanchang 330103,China;National Engineering Technology Research Center for Grum Traditional Chinese Medicine,Dong’e Ejiao Co.,Ltd.,Dong’e 252201,China;College of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,China)

机构地区：[1]济宁医学院药学院,山东日照276826 [2]中国药科大学药学院,江苏南京211198 [3]江西中医药大学药学院,江西南昌330103 [4]东阿阿胶股份有限公司,国家胶类中药工程技术研究中心,山东东阿252201 [5]山东中医药大学药学院,山东济南250355

出　　处：《中成药》2023年第7期2250-2258,共9页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金青年基金项目(81703703)。

摘　　要：目的基于造血活性小肽研究阿胶炮制原理。方法Orbitrap-MS法对阿胶及其炮制品(蛤粉炒、蒲黄炒)的酶解-超滤物进行数据解析,根据分值、丰度、韦恩图、热图聚类分析推断其可能具有造血活性或具有深入研究价值的肽序列。采用Fmoc固相合成法对其进行合成,检测纯度、三氟乙酸残留、净肽含量、内毒素等。以红细胞爆式集落形成单位为指标,检测阿胶及其炮制品的酶解-超滤物和13种合成肽活性。结果共检测出171种肽,其中127个来源于胶原蛋白。蛤粉炒阿胶26种新肽产生,21种肽消失;蒲黄炒阿胶16种新肽产生,52种肽消失,多数肽序列保持不变,但丰度有所改变。以其中13种肽作为对象进行Fmoc合成,发现其均可用于细胞培养。结论阿胶炮制后,造血活性升高,推测是胶原蛋白裂解并经消化产生ejp33(GVVGLPGQR)强活性肽、ejp131(GPAGPSGPPGKDGT)弱活性肽以达到增效作用,且其促造血效果是以ejp10(GPAGPIGPV)、ejp6(GPAGPTGPV)、ejp77(LSSPARSGASL)为主的数种肽的协同作用。AIM To study the processing principles of Asini Corii Colla based on small hematopoietic peptides.METHODS The data of enzymolysis ultrafiltration products of Asini Corii Colla and its processed products(stir-fried with clam powder or cattail)were analyzed by Orbitrap-MS method,and the peptide sequences that may have hematopoietic activity or have in-depth research value were predicted according to the result of score,abundance,Venn diagram and thermographic cluster analysis.The sythesized peptides using Fmoc solid-phase synthesis method had determinations of their purity and net content,residual trifluoroacetic acid,endotoxin,etc.The enzymolysis ultrafiltration products of Asini Corii Colla and its processed products,and the activities of thirteen synthetic peptides underwent detection with burst-forming unit-erythroid as an indicator.RESULTS Among the totally detected one hundred and seventy-one peptides,one hundred and twenty-seven were derivatives from collagen.Asini Corii Colla stir fried with clam powder was observed with twenty-six new peptides,and disappearance of,twenty-one peptides;Asini Corii Colla stir fried with cattail was observed with sixteen new peptides,and disappearance of fifty-two peptides;with most peptide sequences remained unchanged,but the abundance changed.The thirteen peptides selected as targets for Fmoc synthesis can all be used for cell culture.CONCLUSION Processed Asini Corii Colla contributes improved hematopoietic activity.It is speculated that the degraded collagen produces strong active peptide ejp33(GVVGLPGQR)and weak active peptide ejp131(GPAGPSGPPGKDGT)to achieve synergistic effect;and ejp10(GPAGPIGPV),ejp6(GPAGPTGPV)and ejp77(LSSPARSGASL)being the major synergistically hematopoietic peptides.

关 键 词：阿胶 蛤粉炒 蒲黄炒 酶解 Orbitrap-MS 红细胞爆式集落形成单位 Fmoc固相合成 

分 类 号：R282[医药卫生—中药学]