文献类型：期刊文献

英文题名：Extraction and identification of collagen-derived peptides with hematopoietic activity from Colla Corii Asini

作者：Wu, Hongzhong[1,2];Ren, Chunyan[3];Yang, Fang[1,4];Qin, Yufeng[5];Zhang, Yuanxing[4];Liu, Jianwen[1]

机构：[1]E China Univ Sci & Technol, Pharmacol Sch, Shanghai 200237, Peoples R China;[2]HUYA Biosci Int, Shanghai 201203, Peoples R China;[3]Icahn Sch Med Mt Sinai, Dept Struct & Chem Biol, New York, NY 10029 USA;[4]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[5]Shandong Dongeejiao Grp, Jinan 252201, Shandong, Peoples R China

年份：2016

卷号：182

起止页码：129

外文期刊名：JOURNAL OF ETHNOPHARMACOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000373411100014)】；

基金：We are grateful for the financial support from Shandong Dongeejiao Group and the National Nature Science Foundation of China (Project number 20572022).

语种：英文

外文关键词：Colla Corii Asini; E-jiao; Traditional Chinese medicine; Anemia; Hematopoiesis; Cancer

摘要：Ethnopharmacological relevance: Colla Corii Asini is a widely used traditional Chinese medicine to treat anemia with a long history due to its stimulating effect in hematopoiesis, but the components contributing to this effect are still unknown. In this study, we aimed to establish a methodology to isolate the bioactive components and provide pharmacological basis for its usage in treating anemia. Methods: 5-FU and gamma-ray radiation induced anemic mice models were generated by treating with 5-FU at 150 mg/kg body weight and gamma-rays by a 4 MV linear accelerator by total body irradiation using female ICR mice respectively. Oral administration of fraction A was performed by gastric lavage at 1 g/kg and 2 g/kg body weight for 12 days and 25 days and peripheral blood sample was collected from ocular sinus red blood cell (RBC) and white blood cell (WBC) counts every 3 days and 5 days for 5-FU and radiation induced models, respectively. Next, fraction A was separated to A1 and A2 using cation exchange chromatography (IEC) based on ionic strength. Fraction A1 was further separated using reverse phase chromatography (RPC) based on the hydrophobicity first with 0-10% linear gradient, then 20%, 30%, 50% constant gradient of 60% acetonitrile in neutral Na2HPO4 buffer. Peak fractions were pooled, evaporatively dried, and dissolved in ultrapure water. Finally, fraction All was analyzed combining tandem mass spectrometry and proteomic tools and two peptides (peptide 11 and 16) were identified. The hematopoietic effects of multiple fractions and the two peptides were measured using colony-forming units-erythroid (CFU-E), an indication of late erythroid progenitor cells and colony-forming units granulocyte-monocyte (CFU-GM), an indication of granulocyte and monocyte progenitor cells respectively on hematopoietic progenitor cells prepared from bone marrow (Till and Mcculloch 1961). Results: Fraction A at 1 g/kg and 2 g/kg could increase RBC and WBC counts in 5-FU and radiation induced anemic mice models. Fraction A1 at 0.1 mg/ml and 0.5 mg/ml, exhibited stronger hematopoietic activity than fraction A2, both of which were subfractions from fraction A using IEX, by elevated CFU-E and CFU-GM of mouse bone marrow cells. Furthermore, fraction All at 0.1 mg/ml showed stronger CFU-E and CFU-GM than fractions A12 to A14 from RPC separation. Finally, peptide 11 and peptide 16 were identified from tandem mass spectrometry and peptide 11 increased CFU-E and CFU-GM in a dose dependent manner. Conclusions: We combined multiple approaches including chromatography, mass spectrometry, cell based assays, as well as animal studies to identify and demonstrate that the hematopoietic effect of Colla Corii Asini is at least in part from the peptidic components identified using our methodology. This is the first time to isolate peptidic components from Colla Corii Asini, and to provide molecular basis for its usage in treating anemia, which may particularly have the potential to benefit cancer patients suffering from myelosuppression due to radiotherapy or chemotherapy. (C) 2016 Elsevier Ireland Ltd. All rights reserved.