应用生物传感器技术筛选拮抗革兰阴性菌主要病原分子的中药

Application of biosensor technology to screen anti-inflammatory traditional Chinese medicine against CpG DNA and lipid A

目的建立以CpG DNA和脂质A为靶点的生物传感器筛选平台,从传统抗炎中药中筛选同时具有拮抗细菌基因组DNA及脂质A活性的中药并按其活性物质的含量进行排序。方法将CpG DNA和脂质A分别包被于生物传感器的生物素样品池和非衍生板以建立靶点,测定114种中药提取液去鞣质后与CpG DNA的亲和力;选择亲和力较高的35种药物与定量CpG DNA(16.5μg/mL)37℃混合孵育30min,再测定其与CpG DNA的亲和力,以评价中药提取液中活性物质的含量。选择亲和力较高的中药与脂质A反应进行再次筛选。结果 114种中药中,侧柏叶、薄荷、半枝莲等35种中药与CpG DNA具有较高亲和力(>100RU);地骨皮、侧柏叶等14种中药中,与CpG DNA特异性结合的活性物质,测得含量均较高。经另一检测靶点脂质A再次筛选后得到8种特异性亲和力较高的备选中药。结论应用生物传感器跟踪检测技术筛选具有结合革兰阴性菌主要致病分子作用的中药具有可行性。

Objective To apply the biosensor technology to screen anti-inflammatory traditional Chinese medicine（TCM） by tar- get of CpG DNA and lipid A. Then TCM were sorted in accordance with the content of anti-inflammatory efficient materials. Meth- ods CpG DNA and lipid A were immobilized onto the surface of biotin and non-derivatized cuvettes respectively for establishing target. 114 anti inflammatory TCMs were sifted by biosensor through targets of two pathogenic molecules. The quantitative CpG DNA were incubated at 37 ℃ with watery extractive from the TCM having high affinity about 30 min,after that the affinity biosen- sor were used to detect binding force. Lipid A was used for secondary screening. Results Thirty-five TCMs had higher combining competence with CpG DNA,and fourteen species owned more specific binding-CpG DNA compositions. Eight species of TCMs con- tained the active substance with specific affinity to CpG DNA and lipid A. Conclusion It is feasible to screen TCMs by biosensor technology.