Three-dimensional cell bioreactor coupled with high performance liquid chromatography-mass spectrometry for the affinity screening of bioactive components from herb medicine.
Three-dimensional cell bioreactor coupled with high performance liquid chromatography-mass spectrometry for the affinity screening of bioactive components from herb medicine.
复制标题
DOI：
10.1016/j.chroma.2012.04.041
复制
发表时间：
2012-06
期刊：
影响因子：
--
通讯作者：
Zhao-li Mou;Xiao-Ni Qi;Rui-Lin Liu;Jing Zhang;Zhiqi Zhang
中科院分区：
文献类型：
--
作者：
Zhao-li Mou;Xiao-Ni Qi;Rui-Lin Liu;Jing Zhang;Zhiqi Zhang
建立了一种高效、简便的三维细胞生物反应器与高效液相色谱-质谱仪联用的方法，用于中草药多种活性成分的亲和筛选和分析。将癌细胞培养在多孔支架上，形成三维细胞生物反应器。与活细胞和固定细胞相互作用后，比较中药提取物的高效液相指纹图谱，以评价中药成分与细胞的结合特性。选择模型抗癌药物(紫杉醇和白藜芦醇)和非抗癌药物(酮洛芬和青霉素G)进行可行性研究。细胞与药物相互作用30min时，紫杉醇和白藜芦醇(各15μg/ml)的结合度分别为82.2±7.2%和66.1±4.1%，酮洛芬和青霉素G(各15μg/ml)的结合度均小于3%。用该方法对虎杖提取物中的活性成分进行筛选，马兜铃酸A和马兜铃酸B在10μg/ml浓度下的结合度分别为63.0±5.1%和18.8±0.9%。结果表明，该方法具有较高的特异性、高效性和简便性，可用于亲和筛选和分析与细胞相互作用的生物活性成分。
An efficient and convenient method, three-dimensional (3-D) cell bioreactor coupled with high performance liquid chromatography–mass spectrometry was developed for affinity screening and analysis of multiple bioactive components from herbal medicines. Cancer cells were cultured on a porous scaffold to form a 3-D cell bioreactor. After interacting with live and fixed cells, the HPLC fingerprinting chromatograms of herbal medicine extract were compared to evaluate the binding properties of herbal components on cells. Model anticancer drugs (paclitaxel and resveratrol) and non-anticancer drugs (ketoprofen and penicillin G) were chosen to investigate the feasibility. When cell–drug interaction time was 30min, the binding degrees of paclitaxel and resveratrol (each 15μg/ml) were 82.2±7.2% and 66.1±4.1%, and for ketoprofen and penicillin G (each 15μg/ml) were less than 3%. This method was used to screen bioactive components from Polygonum cillinerve (Nakai) Ohwi (PCO) extract, and the binding degrees of two main components in PCO extract (10μg/ml), aristolochic acid A and aristolochic acid B, were 63.0±5.1% and 18.8±0.9%, respectively. These results demonstrated that this method was highly specific, efficient and convenient for affinity screening and analysis of bioactive components interacted with cells.