天然产物研究与开发 ›› 2023, Vol. 35 ›› Issue (12): 2154-2167.doi: 10.16333/j.1001-6880.2023.12.015

• 开发研究 • 上一篇    下一篇

基于UHPLC-QTRAP-MS/MS结合网络药理学探讨不同厂家龟甲胶中氨基酸含量差异及其抗骨质疏松的潜在机制

陈巧巧1,唐   皓2,王   芬1,汤延富1,何清湖3*,余   娜1*   

  1. 1湖南中医药大学药学院,长沙 410208;2湖南省中医药研究院附属医院,长沙 410006;3湖南医药学院,怀化 418000
  • 出版日期:2023-12-28 发布日期:2023-12-27
  • 基金资助:
    湖南中医药大学横向项目(50010190);湖南省研究生科研创新项目(2022CX85);湖南省中医药管理局基金(B2023012);湖南中医药大学大学生创新创业训练计划(1008040)

Difference of amino acid content in Testudinis Carapacis et Plastri Colla from different manufacturers and its potential mechanism of anti-osteoporosis based on UHPLC-QTRAP-MS/MS combined with network pharmacology

CHEN Qiao-qiao1,TANG Hao2,WANG Fen1,TANG Yan-fu1,HE Qing-hu3*,YU Na1*   

  1. 1School of Pharmacy,Hunan University of Traditional Chinese Medicine,Changsha 410208,China;2Hunan Academy of Traditional Chinese Medicine Affiliated Hospoital,Changsha 410006,China;3Hunan Pharmaceutical University,Huaihua 418000,China
  • Online:2023-12-28 Published:2023-12-27

摘要:

比较市面上龟甲胶的质量差异,探索龟甲胶治疗骨质疏松症的潜在活性成分及靶点。我们选取市面上4个厂家共15批次的龟甲胶样品,采用UHPLC-QTRAP-MS/MS检测各龟甲胶中水解氨基酸的含量,多元统计分析各批次中水解氨基酸含量的差异性。采用网络药理学预测龟甲胶的活性成分及治疗骨质疏松症的核心靶点,通过GO与KEGG富集分析预测分子机制,并运用分子对接预测各氨基酸与Hub靶点的结合模式。结果显示,厂家A和D的样品聚类良好,其中A厂家的龟甲胶综合得分最高。17种水解氨基酸中谷氨酸、天冬氨酸、丝氨酸、L-羟脯氨酸、赖氨酸、精氨酸、苏氨酸、蛋氨酸的含量差异对龟甲胶质量影响较大。筛选出17个水解氨基酸的175个潜在靶点,68个骨质疏松症核心靶点,其中甘氨酸、丙氨酸为关键成分。GO/KEGG分析核心靶点主要涉及α-氨基酸代谢过程、细胞氨基酸代谢过程、小分子分解代谢过程等生物进程,以及氨基酸合成、精氨酸和脯氨酸代谢、碳代谢等212条信号通路。分子对接则显示2个关键成分甘氨酸、丙氨酸与11个Hub靶点的活性口袋结合较好。综上,市面上不同厂家及批次间的龟甲胶水解氨基酸含量存在一定差异。龟甲胶抗骨质疏松的关键活性成分可能是甘氨酸、丙氨酸,其作用机理可能与CAT、NOS1、GATM等靶点相关,通过影响氨基酸生物合成、精氨酸和脯氨酸代谢、碳代谢等途径而起作用。

关键词: 龟甲胶, 氨基酸, UHPLC-QTRAP-MS/MS, 骨质疏松症, 多元统计分析, 网络药理学

Abstract:

To compare the quality differences of Testudinis Carapacis et Plastri Colla (TCPC) in the market,and explore the potential active ingredients for osteoporosis,15 batches of TCPC samples from four companies were selected to detect the contents of hydrolysis amino acid by UHPLC-QTRAP-MS/MS.Compared the content differences of hydrolyzed amino acids in 15 batches with multivariate statistics.To predict the active components of TCPC and the core targets for treating osteoporosis with network pharmacology,to predict the molecular mechanism with GO and KEGG enrichment analysis,and the binding mode of amino acid and the Hub target were predicted by molecular docking.The results showed that the samples from companies A and D were well clustered,and the synthesis score of company A was the highest.The contents of glutamate,aspartate,serine,L-hydroxyproline,lysine,arginine,threonine,and methionine had a greater impact on the quality of TCPC.175 potential targets and 68 core targets for treating osteoporosis were selected in the 17 amino acids,of which glycine and alanine were the key components.The core targets mainly involved of the alpha amino acid metabolism,cellular amino acid metabolism,small molecule catabolism and other biological processes,as well as amino acid synthesis,arginine and proline metabolism,carbon metabolism and other 212 signaling pathways.Molecular docking showed that the glycine and alanine were well bound to the active pockets of 11 Hub targets.In conclusion,there are some differences in the amino acid content of TCPC between different companies and batches.The key active ingredients of TCPC anti-osteoporosis may be glycine and alanine,and the mechanism may be related to the targets of CAT,NOS1,GATM,etc.which affecting the pathways of amino acid biosynthesis,arginine and proline metabolism,and carbon metabolism.

Key words:

中图分类号:  R282.74