目的 基于UHPLC-QTOF-MS/MS技术对骆驼蓬Pegannum harmala化学成分进行分析,并探讨其神经保护活性。方法 色谱柱为ACQUITY UPLC BEH C₁₈(100 mm×2.1 mm,1.6 μm),流动相为0.1%甲酸水溶液(A)-0.1%甲酸乙腈溶液(B),梯度洗脱;体积流量0.3 mL/min;进样体积2 μL;柱温35℃。质谱系统采用电喷雾离子源(ESI),正离子模式下检测。利用过氧化氢(H₂O₂)和6-羟基多巴胺(6-hydroxydopamine,6-OHDA)诱导的PC12细胞氧化损伤模型和脂多糖(lipopolysaccharide,LPS)诱导的BV2细胞神经炎症模型,探讨骆驼蓬全草和种子的甲醇提取物以及去氢骆驼蓬碱、骆驼蓬碱、哈尔满和去甲哈尔满4种骆驼蓬中主要生物碱的神经保护活性。结果 从骆驼蓬中共鉴别32个化学成分,全草中主要含有脱氧鸭嘴花碱、鸭嘴花醇等喹唑酮类生物碱,而种子中主要含有去氢骆驼蓬碱、骆驼蓬碱等β-咔啉类生物碱。骆驼蓬全草、种子甲醇提取物和4种生物碱对LPS刺激诱导BV2细胞一氧化氮(NO)生成具有一定的抑制活性,并对H₂O₂和6-OHDA诱导的PC12细胞损伤均具有明显的保护作用(P<0.01)。结论 骆驼蓬全草和种子中的生物碱成分存在差异,而且其所含的生物碱类化合物具有一定的神经保护作用,为骆驼蓬的质量控制及药效物质基础研究提供了参考。

Objective To analyze the chemical constituents of Pegannum harmala based on UHPLC-QTOF-MS/MS and explore its possible neuroprotective activities. Methods The chromatographic column was ACQUITY UPLC BEH C₁₈ (100 mm×2.1mm, 1.6 μm). The mobile phase consisted of 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile solution (B), followed by gradient elution. Volume flow was 0.3 mL/min; The injection volume was 2 μL. Column temperature was 35℃. The mass spectrometry system used electrospray ion source (ESI), which was detected in positive ion mode. Then, the oxidative damage model induced by H₂O₂ and 6-OHDA in PC12 cells and the BV2 neuroinflammation model induced by LPS were established. The extracts of the aerial parts and seeds of P. harmala, and the four main alkaloids of harmine, harmaline, harman and norharman, were investigated on their neuroprotective activities. Results A total of 32 chemical constituents were identified from P. harmala. The aerial parts mainly contained quinazolone alkaloids, such as peganol and deoxypeganine, while the seeds mainly contained β-carboline alkiods, such as harmine and harmaline. The methanol extracts from the aerial parts, seeds and four main alkaloids showed certain inhibitory activity on NO production in BV2 cells stimulated by LPS. They also had obvious protective effects on PC12 cells induced by H₂O₂ and 6-OHDA (P < 0.01). Conclusion The alkaloid components in the aerial parts and the seeds are quite different, and the alkaloid compounds have certain neuroprotective effects. This study provides some reference for the quality control and the basic research of pharmacological substances of P. harmala.

R284.1

国家重点研发计划政府间国际科技创新合作重点专项（2016YFE0129000）；甘肃省重点研发计划（18YF1WA115）；甘肃省自然科学基金项目（20JR5RA311）