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目的:利用H9c2心肌细胞和UHPLC-Q-TOF-MS技术筛选荭草花抗心肌缺血作用的活性成分。方法:用心肌细胞选择性地结合荭草花提取液中的活性成分,洗去未结合的其他成分后,使细胞靶点失活,被结合的活性成分从心肌细胞中释放出来,采用Eclipse Plus C_(18)色谱柱(2.1 mm×100 mm,1.8μm),流动相0.1%甲酸水溶液(A)-0.1%甲酸乙腈溶液(B)梯度洗脱(0~2 min,95%~79%A;2~5 min,79%A;5~6.5 min,79%~60%A;6.5~7.5 min,60%~55%A;7.5~9.5 min,55%~0%A;9.5~10 min,0%~95%A),电喷雾离子源(ESI)正、负离子模式下扫描采集数据,通过采用对照品比对及二级碎片离子对化学成分进行鉴定。结果:检测出荭草花提取物中可与心肌细胞结合的17个成分,采用对照品比对后,确定8个色谱峰所表征的化学成分,依次为原儿茶酸,儿茶素,山柰素-3-O-β-D-葡萄糖苷,槲皮素3-O-α-L-鼠李糖苷,山柰素-3-O-α-L-鼠李糖苷,N-p-香豆酰酪胺,槲皮素和山柰酚。结论:通过分析心肌细胞破碎液中与活性细胞相结合的成分,筛选出了荭草花提取物中能与心肌细胞有相互作用的成分,为明确荭草花的药效物质基础提供了科学依据。
OBJECTIVE: To screen the anti-myocardial ischemic effects of Herba Nuciferae using H9c2 cardiomyocytes and UHPLC-Q-TOF-MS. METHODS: Cardiomyocytes were used to selectively bind the active constituents in the extract of E. purpurea, and after the unbound other components were washed away, the target cells were inactivated and the bound active components were released from the cardiomyocytes. Using Eclipse Plus C_ The mobile phase was eluted with 0.1% formic acid (A) and 0.1% formic acid in acetonitrile (B) with gradient elution (0-2 min, 95% -79% A; 5 to 6.5 min, 79 to 60% A; 6.5 to 7.5 min, 60 to 55% A; 7.5 to 9.5 min, 55 to 0% A; 9.5 to 10 min, 0% ~ 95% A), electrospray ionization (ESI) positive and negative ion mode scan acquisition data, through the use of reference substance and secondary fragment ions to identify the chemical composition. Results: Seventeen components of the extract of Myrtaceae, which could bind to cardiomyocytes, were detected. After comparing with the reference substance, the chemical components of the eight chromatographic peaks were identified, including protocatechuic acid, catechin, 3-O-β-D-glucoside, quercetin 3-O-α-L-rhamnoside, behenin-3-O-α-L-rhamnoside, Np-coumarate Amines, quercetin and kaempferol. CONCLUSION: The constituents that bind to active cells in cardiomyocyte lysate were screened and the components that interact with cardiomyocytes were screened out. The results provided a scientific basis for clarifying the pharmacological substance basis of E. purpurea.