College of Pharmacy, State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, China.
Anal Bioanal Chem. 2012 Nov;404(8):2445-52. doi: 10.1007/s00216-012-6332-9. Epub 2012 Sep 11.
Traditional Chinese medicine (TCM) preparations have been used as an effective multitarget strategy for the treatment of complex diseases; however, their bioactive constituents are undefined and difficult to identify. In this study, a simple and dual-target method based on ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry combined with dual-bioactive (NF-κB and β(2)-adrenergic receptor) luciferase reporter assay systems was developed for the rapid determination of various bioactive compounds of TCM preparations. Qingfei Xiaoyan Wan, a TCM preparation used for the clinical therapy of asthma, was analyzed with this method. Potential anti-inflammatory and spasmolytic constituents were screened using NF-κB and β(2)-adrenergic receptor activity luciferase reporter assay systems and simultaneously identified according to the time-of-flight mass spectrometry data. One β(2)-adrenergic receptor agonist (ephedrine) and four structural types of NF-κB inhibitors (arctigenin derivatives, cholic acid derivatives, chlorogenic acid, and sinapic acid) were characterized. Tracheloside was considered a new NF-κB inhibitor. Further cytokine and chemokine detection confirmed the anti-inflammatory effects of the potential NF-κB inhibitors. The integration of ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry and dual-bioactive human cell functional evaluation systems proved to be a simple and effective strategy for the rapid screening of various bioactive compounds in TCM preparations used to treat complex diseases.
中药制剂一直被用作治疗复杂疾病的有效多靶点策略；然而，其生物活性成分尚未确定，难以识别。在这项研究中，开发了一种基于超高效液相色谱/四极杆飞行时间质谱结合双生物活性（NF-κB 和 β(2)-肾上腺素能受体）荧光素报告检测系统的简单双靶方法，用于快速测定中药制剂的各种生物活性化合物。用这种方法分析了用于哮喘临床治疗的中药制剂清肺消炎丸。使用 NF-κB 和 β(2)-肾上腺素能受体活性荧光素报告检测系统筛选潜在的抗炎和松弛活性成分，并根据飞行时间质谱数据同时进行鉴定。鉴定出一个β(2)-肾上腺素能受体激动剂（麻黄碱）和四种 NF-κB 抑制剂（牛蒡子苷元衍生物、胆酸衍生物、绿原酸和芥子酸）的结构类型。通关藤苷被认为是一种新的 NF-κB 抑制剂。进一步的细胞因子和趋化因子检测证实了潜在 NF-κB 抑制剂的抗炎作用。超高效液相色谱/四极杆飞行时间质谱与双生物活性人细胞功能评价系统的结合，被证明是一种快速筛选用于治疗复杂疾病的中药制剂中各种生物活性化合物的简单有效策略。
