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中国临床药理学与治疗学 ›› 2024, Vol. 29 ›› Issue (9): 988-1001.doi: 10.12092/j.issn.1009-2501.2024.09.004

• 基础研究 • 上一篇    下一篇

基于代谢组学与网络药理学研究四黑方对斑马鱼黑色素的作用机制

苏琪辉1,王婧1,雒荣荣1,黄玉荣1,李欣1,王颖莉1,贾颖2   

  1. 1山西中医药大学,中药与食品工程学院,晋中  030619,山西;2山西中医药大学,第一临床学院,太原  030024,山西

  • 收稿日期:2023-12-25 修回日期:2023-11-18 出版日期:2024-09-26 发布日期:2024-08-21
  • 通讯作者: 王颖莉,女,博士,教授,研究方向:中药药效及物质基础研究。 E-mail:wyltyut@163.com
  • 作者简介:苏琪辉,女,硕士研究生,研究方向:中药新产品开发与应用。 E-mail:suqihui@stu.sxtcm.edu.cn
  • 基金资助:
    中药外用新制剂工艺开发研究;山西省中医药管理局科研课题;山西省卫生健康委科研课题

Study on the mechanism of action of Siheifang on zebrafish melanin based on metabolomics and network pharmacology

SU Qihui1, WANG Jing1, LUO Rongrong1, HUANG Yurong1, LI Xin1, WANG Yingli1, JIA Ying2   

  1. 1Shanxi University of Chinese Medicine, College of Traditional Chinese Medicine and Food Engineering, Jinzhong 030619, Shanxi, China; 2Shanxi University of Chinese Medicine, First Clinical College, Taiyuan 030024, Shanxi, China
  • Received:2023-12-25 Revised:2023-11-18 Online:2024-09-26 Published:2024-08-21
  • Supported by:
    中药外用新制剂工艺开发研究(2022SYS23);山西省卫生健康委科研课题(2020096);山西省中医药管理局科研课题(2022ZYYC098)

摘要:

目的:联合网络药理学与代谢组学研究四黑方(SHF)改善色素缺失症(pigment deficiency disease,PD)的作用机制。方法:以1-苯基-2-硫脲(phenylthiourea,PTU)诱导的色素缺失性疾病斑马鱼胚胎为动物模型,分析SHF提取物(0.01、0.02、0.04 mg/mL)对斑马鱼胚胎黑色素细胞形态、黑色素面积、酪氨酸酶活性、黑色素含量的影响;采用超高效液相色谱-质谱(ultra high performance liquid chromatography-mass spectrometry,UHPLC-MS)对SHF治疗缺黑色素斑马鱼胚胎模型筛选差异代谢物,获得相关代谢通路。运用网络药理学获取SHF治疗PD的关键靶点,进行KEGG通路富集分析。将鉴定得到的差异代谢物和SHF与PD交集靶点导入Metscape插件,建立“代谢物-反应-酶-基因”网络,寻找关键代谢物、靶点和代谢通路。结果:SHF治疗后能增加斑马鱼黑色素形成,激活酪氨酸酶活性,提高黑色素含量。代谢组学分析获得54个差异代谢物,对上述代谢物进行代谢通路分析,涉及苯丙氨酸、酪氨酸和色氨酸的生物合成、甘油磷脂代谢、酪氨酸代谢、亚油酸代谢、氨酰基-tRNA生物合成等通路。网络药理学得到成分和疾病交集靶点55个,KEGG涉及胰腺癌、TNF、癌症等信号通路。代谢组学和网络药理学联合分析确定了4个关键靶点:COMT、CYP1B1、TYR、ALDH2;3个关键代谢物:L-Tyrosine、homovanllate、L-Lysine;3个重要代谢途径:酪氨酸代谢、缬氨酸/亮氨酸/异亮氨酸降解、赖氨酸代谢。结论:SHF对PD具有良好的改善作用,联合代谢组学与网络药理学得到SHF可能通过调控L-Tyrosine这一代谢物,增强对酪氨酸代谢通路的影响,从而促进黑色素的形成。

关键词: 四黑方, 色素缺失症, 代谢组学, 网络药理学

Abstract:

AIM: To study the mechanism of Siheifang (SHF) in improving pigment deficiency disease (PD) by combining network pharmacology and metabolomics. METHODS: Using zebrafish embryos with pigment deficiency disease induced by 1-phenyl-2-thiourea (PTU) as an animal model, the effects of SHF extract (0.01, 0.02, 0.04 mg/mL) on the morphology, melanin area, tyrosinase activity, and melanin content of zebrafish embryos were analyzed. Ultra high performance liquid chromatography-mass spectrometry (UHPLC-MS) was used to screen differential metabolites and obtain relevant metabolic pathways in the SHF treatment of melanin deficient zebrafish embryos model. Network pharmacology was used to obtain key targets for SHF treatment of PD and conduct KEGG pathway enrichment analysis. Import The identified differential metabolites and SHF PD intersection targets were imported into the Metscape plugin, to establish a "metabolite reaction enzyme gene" network, and search for key metabolites, targets, and metabolic pathways. RESULTS: SHF treatment could increase the formation of zebrafish melanin, activate tyrosinase activity, and increase melanin content. Metabolomics analysis obtained 54 differential metabolites, and metabolic pathway analysis was conducted on these metabolites, involving the biosynthesis of phenylalanine, tyrosine, and tryptophan, glycerol phospholipid metabolism, tyrosine metabolism, linoleic acid metabolism, and aminoacyl tRNA biosynthesis pathways. Network pharmacology had obtained 55 cross targets of components and diseases. KEGG involved pancreatic cancer, TNF, cancer and other signal pathways. The joint analysis of metabolomics and network pharmacology identified four key targets: COMT, CYP1B1, TYR, and ALDH2; three key metabolites: L-tyrosine, homovanllate, L-lysine; three important metabolic pathways: tyrosine metabolism, valine/leucine/isoleucine degradation, and lysine metabolism. CONCLUSION: SHF has a good improvement effect on PD, and combined with metabolomics and network pharmacology, SHF may enhance its influence on the tyrosine metabolism pathway by regulating the metabolite L-tyrosine, thereby promoting the formation of melanin.

Key words: Siheifang, pigmentation deficiency, metabolomics, network pharmacology

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