細胞膜色譜在線HPLC-IT-TOF MS聯(lián)用系統(tǒng)篩選分析射干中次野鳶尾黃素的抗EGFR活性作用
發(fā)布時間：2019-01-11 08:00
【摘要】：中藥射干具有多種藥理作用并得到廣泛研究,但對其抗腫瘤活性組分的研究較少。本研究建立了一種用于篩選射干中抗腫瘤活性組分的方法。利用高表達表皮生長因子受體(EGFR)細胞制備細胞膜色譜柱,并通過十通切換閥與高效液相色譜-離子阱-飛行時間質(zhì)譜(HPLC-IT-TOF MS)構(gòu)建成二維在線聯(lián)用系統(tǒng)。利用吉非替尼驗證該系統(tǒng)的適用性,分子模擬對接實驗研究活性組分與受體的相互作用方式,并利用MTT實驗研究所篩選組分對EGFR細胞的體外抑制作用。結(jié)果表明,利用該系統(tǒng)可從射干總提取物中篩選出能夠作用于EGFR的活性成分,并鑒定為次野鳶尾黃素;次野鳶尾黃素與EGFR的相互作用方式與陽性藥吉非替尼相似;在濃度為0.4~50μmol/L范圍內(nèi),吉非替尼、次野鳶尾黃素對高表達EGFR細胞的體外抑制作用具有劑量依賴性。EGFR細胞膜色譜在線HPLC-IT-TOF MS法有望成為從中草藥中發(fā)現(xiàn)潛在抗腫瘤候選藥物的有效方法。
[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.
【作者單位】： 西安交通大學藥學院;
【基金】：國家自然科學基金(81503032、81230079、81227802) 國家博士后基金項目(2015M570844、2016T90930) 陜西省博士后科學基金 中央高校基本科研業(yè)務(wù)費資助
【分類號】：O657.63;R285
本文編號：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.
【作者單位】： 西安交通大學藥學院;
【基金】：國家自然科學基金(81503032、81230079、81227802) 國家博士后基金項目(2015M570844、2016T90930) 陜西省博士后科學基金 中央高校基本科研業(yè)務(wù)費資助
【分類號】：O657.63;R285
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