中国中药杂志

2021, v.46(18) 4833-4840

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UPLC-MS/MS研究荭草提取物的H9c2细胞药代动力学
Pharmacokinetic study of Polygonum orientale extract in H9c2 cells by UPLC-MS/MS

陆苑;黎娜;吕婷;王永林;潘洁;孙佳;李勇军;刘春花;
LU Yuan;LI Na;LYU Ting;WANG Yong-lin;PAN Jie;SUN Jia;LI Yong-jun;LIU Chun-hua;Provincial Key Laboratory of Pharmaceutics in Guizhou Province, State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Medical University;School of Pharmaceutical Sciences, Guizhou Medical University;Engineering Research Center for the Development and Application of Ethnic Medicine and TCM (Ministry of Education), Guizhou Medical University;

摘要(Abstract):

建立H9c2细胞中异荭草素、荭草素、槲皮苷、牡荆素和山柰素-3-O-β-D-葡萄糖苷的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法,并运用在荭草提取物的H9c2细胞药代动力学研究中。荭草提取物(100μg·mL~(-1))处理H9c2细胞后,于不同时间点分别收集获得整体细胞以及相应的细胞核、线粒体和高尔基体,蛋白沉淀方法处理样品,采用UPLC-MS/MS测定整体细胞以及亚细胞结构中异荭草素、荭草素、槲皮苷、牡荆素、山柰素-3-O-β-D-葡萄糖苷的药物浓度,并计算相关药动学参数。结果在整体细胞中,各成分的达峰时间均为8 h,荭草素、牡荆素、槲皮苷、异荭草素与细胞核和线粒体的亲和力均较高,山柰素-3-O-β-D-葡萄糖苷与线粒体亲和力较高。实验结果提示荭草保护心肌细胞中,这5个化学成分可能主要是通过作用于细胞核或线粒体而发挥药理效应。
A detection method of ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was established to detect concentrations of isoorientin, orientin, quercetin, vitexin and kaempferol-3-O-β-D-glucoside in H9 c2 cells and applied to the pharmacokinetic study of Polygonum orientale extract in the cells. H9 c2 cells were treated with 100 μg·mL~(-1) P. orientale extract and then they and the corresponding nuclei, mitochondria and Golgi bodies were collected at the set time. After protein precipitation, UPLC-MS/MS was used to determine concentrations of isoorientin, orientin, quercetin, vitexin and kaempferol-3-O-β-D-glucoside in the whole cells and subcellular structures. Also, related pharmacokinetic parameters were calculated. The results showed that the peak time was 8 h for all these components. Orientin, vitexin, quercetin and isoorientin have high affinities to nuclei and mitochondria, while the affinity of kaempferol-3-O-β-D-glucoside is higher with mitochondria compared to nuclei. It is suggested that these chemical components of P. orientale may mainly act on nuclei or mitochondria to exert pharmacological effects of protecting cardiomyocytes.

关键词(KeyWords): UPLC-MS/MS;荭草;H9c2细胞;细胞药代动力学;细胞核;线粒体
UPLC-MS/MS;Polygonum orientale;H9c2 cell;cellular pharmacokinetics;nuclei;mitochondria

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81803700,U1812403-05,81660712);; 贵州省自然科学基金项目(黔科合基础[2020]1Z068);; 中央引导地方科技专项项目(黔科中引地[2018]4006);; 贵州省科学技术厅人才团队项目(黔科合平台人才[2016]5613/677);; 贵州医科大学学术新苗培养及创新探索专项(黔科合平台[2017]5718)

作者(Author): 陆苑;黎娜;吕婷;王永林;潘洁;孙佳;李勇军;刘春花;
LU Yuan;LI Na;LYU Ting;WANG Yong-lin;PAN Jie;SUN Jia;LI Yong-jun;LIU Chun-hua;Provincial Key Laboratory of Pharmaceutics in Guizhou Province, State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Medical University;School of Pharmaceutical Sciences, Guizhou Medical University;Engineering Research Center for the Development and Application of Ethnic Medicine and TCM (Ministry of Education), Guizhou Medical University;

Email:

DOI: 10.19540/j.cnki.cjcmm.20210618.202

参考文献(References):

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