基于液质联用的川芎嗪对人THP-1细胞干预作用的代谢组学研究
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摘要
目的采用代谢轮廓及代谢通路技术靶向性分析人白血病THP-1细胞的代谢物,研究白血病发病机制及川芎嗪的干预作用。方法采用超高效液相-四极杆静电场轨道肼质谱技术(UHPLC-Q-Exactive)靶向性分析人THP-1细胞代谢物的含量变化及的代谢途径变化,通过聚焦分析方法明确白血病发病的核心靶标及相关代谢径路。同时,观察给予川芎嗪对人THP-1细胞在代谢调控方面的治疗作用。结果通过化学计量学综合分析显示,THP-1细胞存在显著的代谢异常。差异代谢物分别为花生四烯酸、柠檬酸、肌酸、丙酮酸盐、胆碱、苯基丙氨酸。给予川芎嗪后其含量全部呈现回调趋势。结论川芎嗪单体对人白血病THP-1细胞有很好的抑制作用,通过靶向性的分析人THP-1细胞发现,其与正常细胞相比主要存在显著的代谢差异,所主要涉及的花生四烯酸代谢、丙酮酸代谢、苯丙氨酸代谢、柠檬酸循环(TCA循环)等通路可是为人白血病THP-1细胞的核心代谢途径,本研究鉴定的差异代谢物对急性髓系白血病的诊断和治疗具有重要意义,为进一步研究川芎嗪治疗急性髓系白血病的作用机制提供理论依据。
Objective To analyze the metabolites of THP-1 cells in human leukemia by metabolic profiling and metabolic pathway technology, and revealed the pathogenesis of leukemia and the intervention effect of ligustrazine. Methods Ultra-high performance liquid chromatography-quadrupole electrostatic field orbital hydrazine mass spectrometry(UHPLC-Q-Exactive) was used to analyze the changes of metabolites and metabolic pathways in human THP-1 cells. The core targets and related metabolic pathways of leukemia were identified by focusing analysis. Meanwhile, the therapeutic effects of ligustrazine on THP-1 cells in metabolic regulation were observed. Results Chemometrics analysis showed that THP-1 cells had significant metabolic abnormalities. The differential metabolites were arachidonic acid, citric acid, creatine, pyruvate, choline and phenylalanine. After giving ligustrazine, the content of ligustrazine showed a trend of regression. Conclusion Ligustrazine monomer has a good inhibitory effect on human leukemia THP-1 cells. By targeting analysis, human THP-1 cells are found to have significant metabolic differences compared with normal cells. Arachidonic acid metabolism, pyruvate metabolism, phenylalanine metabolism, citric acid cycle and other pathways are the core generation of human leukemia THP-1 cells. The differential metabolites identified in this study have important significance in the diagnosis and treatment of acute myeloid leukemia, and provide theoretical basis for further study of the mechanism of ligustrazine in the treatment of acute myeloid leukemia.
Objective To analyze the metabolites of THP-1 cells in human leukemia by metabolic profiling and metabolic pathway technology, and revealed the pathogenesis of leukemia and the intervention effect of ligustrazine. Methods Ultra-high performance liquid chromatography-quadrupole electrostatic field orbital hydrazine mass spectrometry(UHPLC-Q-Exactive) was used to analyze the changes of metabolites and metabolic pathways in human THP-1 cells. The core targets and related metabolic pathways of leukemia were identified by focusing analysis. Meanwhile, the therapeutic effects of ligustrazine on THP-1 cells in metabolic regulation were observed. Results Chemometrics analysis showed that THP-1 cells had significant metabolic abnormalities. The differential metabolites were arachidonic acid, citric acid, creatine, pyruvate, choline and phenylalanine. After giving ligustrazine, the content of ligustrazine showed a trend of regression. Conclusion Ligustrazine monomer has a good inhibitory effect on human leukemia THP-1 cells. By targeting analysis, human THP-1 cells are found to have significant metabolic differences compared with normal cells. Arachidonic acid metabolism, pyruvate metabolism, phenylalanine metabolism, citric acid cycle and other pathways are the core generation of human leukemia THP-1 cells. The differential metabolites identified in this study have important significance in the diagnosis and treatment of acute myeloid leukemia, and provide theoretical basis for further study of the mechanism of ligustrazine in the treatment of acute myeloid leukemia.
引文
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[11]梁蓉,杨平地,陈协群.川芎嗪和异博定联合逆转HL60/HT细胞的多药耐药[J].第四军医大学学报,1998(4):385-387.
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[2]都本敏,荆进红.川芎嗪的临床应用[J].现代中西医结合杂志,2006,15(10):1398-1399.
[3]徐浩.川芎嗪的临床应用和药理作用--川芎嗪心血管药理与钙拮抗作用[J].中国中西医结合杂志,2003,23(5):376-377.
[4]梁爱群.川芎嗪的药理及机理研究[J].时珍国医国药,2005,16(6):532-533.
[5]伍俞霓.川芎嗪对HL-60细胞株及其SCID小鼠模型的治疗机制研究[D].重庆:重庆医科大学,2012.
[6]伍俞霓,徐酉华,顾晓艳,等.川芎嗪诱导HL-60细胞株分化与分子机制的研究[J].中国中药杂志,2011,36(21):3007-3011.
[7]范可,李中明.川芎嗪影响血管内皮生长因子诱导HL-60白血病细胞增殖的效应(英文)[J].中国组织工程研究与临床康复,2008(16):3163-3166.
[8]梁蓉,杨平地,陈协群.川芎嗪对白血病细胞株HL-60和K562的生长抑制反应[J].第四军医大学学报,1998(1):109-110.
[9]汪明宇.川芎嗪对HL-60细胞增殖和侵袭能力的影响的研究[D].重庆:重庆医科大学,2007.
[10]梁蓉,杨平地,陈协群,等.川芎嗪对白血病HL-60/VCR细胞多药耐药的逆转及其机制研究[J].中华血液学杂志,1999(6):42-43.
[11]梁蓉,杨平地,陈协群.川芎嗪和异博定联合逆转HL60/HT细胞的多药耐药[J].第四军医大学学报,1998(4):385-387.
[12]梁蓉,杨平地,陈协群.川芎嗪和(或)环孢素A对HL60/HT细胞耐药的逆转[J].中华内科杂志,1999(4):43-44.
[13]梁蓉,陈协群,杨平地.川芎嗪对HL60/HT耐药细胞多种化疗药物的增敏作用[J].中国中西医结合杂志,2001(S1):130-132.