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 Guiyang 550004, China.
Provincial Key Laboratory of Pharmaceutics in Guizhou Province, State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Medical University Guiyang 550004, China School of Pharmaceutical Sciences, Guizhou Medical University Guiyang 550004, China.
Zhongguo Zhong Yao Za Zhi. 2021 Sep;46(18):4833-4840. doi: 10.19540/j.cnki.cjcmm.20210618.202.
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.
建立了超高效液相色谱-串联质谱法（UPLC-MS/MS）检测H9 c2细胞中异荭草苷、荭草苷、槲皮素、牡荆素和山柰酚-3-O-β-D-葡萄糖苷的浓度，并将其应用于蓼蓝提取物在细胞中的药代动力学研究。用100 μg·mL⁻¹蓼蓝提取物处理H9 c2细胞，然后在设定时间收集细胞及其相应的细胞核、线粒体和高尔基体。经过蛋白沉淀后，采用UPLC-MS/MS测定全细胞和亚细胞结构中异荭草苷、荭草苷、槲皮素、牡荆素和山柰酚-3-O-β-D-葡萄糖苷的浓度。同时，计算相关药代动力学参数。结果表明，所有这些成分的峰时间均为8小时。荭草苷、牡荆素、槲皮素和异荭草苷对细胞核和线粒体具有较高的亲和力，而山柰酚-3-O-β-D-葡萄糖苷与线粒体的亲和力高于细胞核。提示蓼蓝的这些化学成分可能主要作用于细胞核或线粒体，发挥保护心肌细胞的药理作用。
