建立启膈方UPLC指纹图谱并验证启膈方对食管癌转移的影响  

Establishment of UPLC Fingerprint of Qige Formula and Verification of Its Effect on Metastasis of Esophageal Cancer

作　　者：史会娟[1] 石冬璇 孙亚琴 于富洋 吴忠冰 张玉双[1] 李晶[1,2] SHI Hui-juan;SHI Dong-xuan;SUN Ya-qin;YU Fu-yang;WU Zhong-bing;ZHANG Yu-shuang;LI Jing(The Fourth Hospital of Hebei Medical University,Shijiazhuang 050200,China;College of Integrated Chinese and Western Medicine of Hebei Medical University,Shijiazhuang 050200,China;Shijiazhuang Yiling Pharmaceutical Co.,Ltd,Shijiazhuang 050200,China)

机构地区：[1]河北医科大学第四医院,石家庄050200 [2]河北医科大学中西医结合学院,石家庄050200 [3]石家庄以岭药业股份有限公司,石家庄050200

出　　处：《中国中医基础医学杂志》2022年第4期619-624,共6页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然基金面上项目(81973761)-甘润濡养法改善食管癌微环境中巨噬细胞极化调控Gas6/Axl信号通路抑制转移的研究;河北省科技厅河北省重点研发计划项目(192777115D)-四物饮调控血管内皮生长因子抑制局部晚期食管癌复发的研究。

摘　　要：目的:观察启膈方对食管癌细胞迁移及侵袭的抑制作用,并建立启膈方的超高效液相(ultra performance liquid chromatography,UPLC)指纹图谱。方法:制备启膈方冻干粉,将高分化食管癌细胞Eca109分成对照组和启膈方组,应用划痕实验检测经启膈方干预前后食管癌细胞迁移能力的改变,借助荧光显微镜观察启膈方干预前后细胞形态的变化,采用蛋白芯片观察启膈方干预后肿瘤相关生长因子的变化,采用ACQUITY CSH C18(2.1mm×100 mm,1.7μm)色谱柱,梯度洗脱、流速0.4 mL/min、检测波长254 nm、柱温40℃,并建立启膈方冻干粉UPLC指纹图谱。结果:启膈方组细胞迁移能力明显低于对照组(P<0.05),荧光显微镜观察显示经启膈方干预后细胞形态规则,蛋白芯片结果显示启膈方能够降低肿瘤相关生长因子的表达,10批启隔方冻干粉相似度均在0.9以上,共标定了17个共有峰,指认出7个色谱峰,分别为迷迭香酸、异槲皮苷、丹酚酸A、丹酚酸B、丹酚酸E、金丝桃苷、紫草酸。结论:启膈方能够降低肿瘤相关生长因子的蛋白表达,从而降低食管癌细胞迁移能力,建立的启膈方冻干粉UPLC指纹图谱可为启膈方的质量评价及药效物质基础研究提供依据和参考。Objective:To observe the inhibitory effect of Qige formula on the migration and invasion of esophageal cancer cells,and to establish the ultra performance liquid chromatography(UPLC)fingerprint of Qige formula.Methods:The water extract lyophilized powder of Qige formula was prepared,and Eca109 cells were divided into the control group and Qige formula group.Scratch test was used to detect the migration ability of esophageal cancer cells after the intervention of Qige formula.Fluorescence microscope was used to observe the changes of cell morphology before and after the intervention.Protein chip was used to observe the changes of tumor-related growth factors after the intervention.The chromatography fingerprint was obtained with ACQUITY CSH C_(18)(2.1mm×100 mm,1.7μm)and gradient elute with flow rate 0.4 mL/min.The detected wavelength was 254 nm,and the column temperature was set at 40℃.Results:The cell migration ability of the Qige formula group was significantly suppressed compared with that of the control group(P<0.05).Fluorescence microscope observation showed that the cell morphology was regulated after the intervention of Qige formula.Protein chip results showed that Qige formula could reduce the expression of tumor-related growth factor.The common pattern of freeze-dried powder fingerprint of Qige formula was produced.The similarity of ten batches of Qige formula lyophilized powder was above 0.9.A total of 17 common peaks were calibrated and 7 chromatographic peaks were identified,namely rosmarinic acid,isoquercetin,salvianolic acid A,salvianolic acid B,salvianolic acid E,hypericin and zioxalic acid.Conclusion:Qige formula can reduce the protein expression of tumor-associated growth factors and reduce the migration ability of esophageal cancer cells.The established UPLC fingerprint of Qige formula freeze-dried powder can provide basis and reference for quality evaluation and pharmacodynamic material basis research of Qige formula.

关 键 词：启膈方 食管癌 细胞迁移 指纹图谱 

分 类 号：R284.1[医药卫生—中药学] R917[医药卫生—中医学]