细胞膜色谱在线HPLC-IT-TOF MS联用系统筛选分析射干中次野鸢尾黄素的抗EGFR活性作用
发布时间：2019-01-11 08:00
【摘要】：中药射干具有多种药理作用并得到广泛研究,但对其抗肿瘤活性组分的研究较少。本研究建立了一种用于筛选射干中抗肿瘤活性组分的方法。利用高表达表皮生长因子受体(EGFR)细胞制备细胞膜色谱柱,并通过十通切换阀与高效液相色谱-离子阱-飞行时间质谱(HPLC-IT-TOF MS)构建成二维在线联用系统。利用吉非替尼验证该系统的适用性,分子模拟对接实验研究活性组分与受体的相互作用方式,并利用MTT实验研究所筛选组分对EGFR细胞的体外抑制作用。结果表明,利用该系统可从射干总提取物中筛选出能够作用于EGFR的活性成分,并鉴定为次野鸢尾黄素;次野鸢尾黄素与EGFR的相互作用方式与阳性药吉非替尼相似;在浓度为0.4~50μmol/L范围内,吉非替尼、次野鸢尾黄素对高表达EGFR细胞的体外抑制作用具有剂量依赖性。EGFR细胞膜色谱在线HPLC-IT-TOF MS法有望成为从中草药中发现潜在抗肿瘤候选药物的有效方法。
[Abstract]:Phytophthora chinensis has a variety of pharmacological effects and has been widely studied, but its antitumor active components are less studied. In this study, a method for screening anti-tumor active components in perforated stem was established. Cell membrane chromatographic column was prepared from (EGFR) cells with high expression of epidermal growth factor receptor (EGF), and a two-dimensional on-line system was constructed by high performance liquid chromatography-ion trap-time-of-flight mass spectrometry (HPLC-IT-TOF MS). Gifitinib was used to verify the applicability of the system, molecular simulation and docking experiments were used to study the interaction between active components and receptors, and MTT was used to screen the inhibitory effects of components on EGFR cells in vitro. The results showed that the active components which could act on EGFR could be isolated from the total extract of Radix Sheidan, and identified as Iris asiaticum, and the interaction between EGFR and EGFR was similar to that of the positive drug Gifitinib. In the concentration range of 0.4 渭 mol/L, gefitinib, The inhibition of irisin on overexpression of EGFR cells in vitro is dose-dependent, and EGFR cell membrane chromatographic on-line HPLC-IT-TOF MS method is expected to be an effective method for the discovery of potential antitumor drugs from Chinese herbal medicine.
【作者单位】： 西安交通大学药学院;
【基金】：国家自然科学基金(81503032、81230079、81227802) 国家博士后基金项目(2015M570844、2016T90930) 陕西省博士后科学基金 中央高校基本科研业务费资助
【分类号】：O657.63;R285
本文编号：2406881
[Abstract]:Phytophthora chinensis has a variety of pharmacological effects and has been widely studied, but its antitumor active components are less studied. In this study, a method for screening anti-tumor active components in perforated stem was established. Cell membrane chromatographic column was prepared from (EGFR) cells with high expression of epidermal growth factor receptor (EGF), and a two-dimensional on-line system was constructed by high performance liquid chromatography-ion trap-time-of-flight mass spectrometry (HPLC-IT-TOF MS). Gifitinib was used to verify the applicability of the system, molecular simulation and docking experiments were used to study the interaction between active components and receptors, and MTT was used to screen the inhibitory effects of components on EGFR cells in vitro. The results showed that the active components which could act on EGFR could be isolated from the total extract of Radix Sheidan, and identified as Iris asiaticum, and the interaction between EGFR and EGFR was similar to that of the positive drug Gifitinib. In the concentration range of 0.4 渭 mol/L, gefitinib, The inhibition of irisin on overexpression of EGFR cells in vitro is dose-dependent, and EGFR cell membrane chromatographic on-line HPLC-IT-TOF MS method is expected to be an effective method for the discovery of potential antitumor drugs from Chinese herbal medicine.
【作者单位】： 西安交通大学药学院;
【基金】：国家自然科学基金(81503032、81230079、81227802) 国家博士后基金项目(2015M570844、2016T90930) 陕西省博士后科学基金 中央高校基本科研业务费资助
【分类号】：O657.63;R285
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