基于UPLC-Q-orbitrap-MS技术探究西黄丸抗乳腺增生的作用机制

Exploring the mechanism of Xihuang Pill's anti-hyperplasia of mammary glands effect based on UPLC-Q-orbitrap-MS technology

基于UPLC-Q-orbitrap-MS与生物网络分析工具系统分析西黄丸改善乳腺组织增生的作用机制。肌注苯甲酸雌二醇和黄体酮建立乳腺增生大鼠模型,采用LC-MS的组织代谢组学探索西黄丸改善乳腺组织增生的关键代谢物和代谢途径。整合生物网络分析工具对西黄丸调节的关键代谢物进行网络分析,聚焦关键代谢通路,挖掘西黄丸改善乳腺组织增生的潜在靶点。与空白组比,模型组大鼠组织中有49种差异代谢物含量有显著差异(P<0.05),西黄丸能显著回调L-丙氨酸、苏氨酸、吲哚-3-羧酸醛、赖氨酸、精氨酸、丙氨酰亮氨酸、甘氨酰络氨酸、γ-谷氨酰亮氨酸、维生素B3、丝氨酰亮氨酸、苏氨酰亮氨酸、异亮氨酰谷氨酸、γ-谷氨酰酪氨酸、癸酰-L-肉碱、2,6,8-三羟基嘌呤、亮氨酰亮氨酸、S-腺苷-蛋氨酸等17种代谢物。对西黄丸调控的关键代谢物进一步网络分析及文献研究表明,晚期糖基化终末产物(AGE)-晚期糖基化终产物受体(RAGE)信号通路可能是西黄丸改善乳腺组织增生的重要通路之一,AGE-RAGE信号通路上的STAT3、MAPK1、EGFR、CASP3、CASP8、PRKCA、JUN等7个蛋白可能为西黄丸改善乳腺组织增生的潜在作用靶点。本文涉及的动物实验操作均遵循甘肃中医药大学动物伦理委员会的规定并通过动物实验伦理审查(批号:2022-705)。

Based on UPLC-Q-orbitrap-MS and biological network analysis tools,the mechanism of Xihuang Pill in improving hyperplasia of mammary glands was systematically analyzed.The rat model of hyperplasia of mammary glands was established by intramuscular injection of estradiol benzoate and progesterone.LC-MS tissue metabolomics was used to explore the key metabolites and metabolic pathways of Xihuang Pill in improving hyperplasia of mammary glands in rat.The network analysis of the key metabolites regulated by Xihuang Pill was carried out by integrating biological network analysis tools,focusing on the key metabolic pathways,and exploring the potential targets of Xihuang Pill to improve hyperplasia of mammary glands.Compared with the control group,there were significant differences in the content of 49 differential metabolites in the tissues of the model group(P<0.05).Xihuang Pills could significantly call back 17 metabolites such as L-alanine,threonine,indole-3-carboxylic aldehyde,lysine,arginine,alanylleucine,glycyltyrosine,γ-glutamyl leucine,vitamin B3,serine leucine,threonine leucine,isoleucine glutamic acid,γ-glutamyl tyrosine,decanoyl-L-carnitine,uric acid,leucylleucine,S-adenosylmethionine.Further network analysis and literature research on the key metabolites regulated by Xihuang Pills showed that the AGE-RAGE signaling pathway may be one of the important pathways for Xihuang Pills to improve hyperplasia of mammary glands.STAT3,MAPK1,EGFR,CASP3,CASP8,PRKCA and JUN in the AGERAGE signaling pathway may be potential targets for Xihuang Pills to improve hyperplasia of mammary glands.The animal experiment operations involved in this paper follow the provisions of the Animal Ethics Committee of Gansu University of Traditional Chinese Medicine and pass the ethical review of animal experiments(approval number:2022-705).