柔性水凝胶细胞筛药基底评估清热凉血中药复方的抗菌效果  

Evaluation of antibacterial effect of heat-clearing and blood-cooling Chinese medicine compounds using flexible hydrogel cell-substrate for drug-screening

作　　者：吕若一 王潇 穆君宇 司晓慧 孙志刚 刘晓晔[1,2] LYU Ruoyi;WANG Xiao;MU Junyu;SI Xiaohui;SUN Zhigang;LIU Xiaoye(Beijing Traditional Chinese Veterinary Engineering Center and Beijing Key Laboratory of Traditional Chinese Veterinary Medicine,Beijing University of Agriculture,Changping,Beijing 102206,China;Animal Science and Technology College,Beijing University of Agriculture,Changping,Beijing 102206,China)

机构地区：[1]北京农学院、兽医学(中医药)、北京市重点实验室、北京市中兽药工程技术研究中心,北京昌平102206 [2]北京农学院动物科学技术学院,北京昌平102206

出　　处：《中国兽医学报》2024年第3期587-594,共8页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(32202864);北京市自然科学基金资助项目(6224060);北京农学院青年教师科研创新能力提升计划资助项目(QJKC2022028);北京农学院科技创新“火花行动”支持计划资助项目(BUA-HHXD2022007);北京市教育委员会科研计划资助项目(KM202310020006)。

摘　　要：抗菌药物的筛选技术对于兽医临床合理精准制定抗菌策略至关重要,传统的筛药技术较多关注药物和病原菌、细胞之间的相互作用,没有考虑到宿主体内微环境对药物、细菌和细胞的影响,所以在筛选具有多靶点宿主导向特点的中药复方时参考性较低。用柔性水凝胶在体外重塑细胞生存的微环境模型,筛选4种经典多靶点中药复方的抗菌效果。利用聚丙烯酰胺凝胶、结合微图案复刻的方法,建立类组织细胞微环境的水凝胶,在水凝胶上形成细胞培养芯片,建立金黄色葡萄球菌侵染细胞的感染互作模型,之后加入中药复方治疗。通过荧光技术可视化检测细胞、细菌的数量变化,实现对中药复方可视化和定量化的药效评价。结果显示,牛角地黄汤、清营汤和清瘟败毒饮均有显著的抗菌作用,其中清瘟败毒饮的抗菌效果最佳且对细胞损伤较小,而白头翁汤对细胞毒性较大且抗菌作用不显著。The screening technology of antibacterial drugs is critical for the rational and accurate formulation of antibacterial strategies in veterinary clinic.However,the screening technology that only focuses on the interaction between drugs and bacteria is not enough to screen multi-target host-directed antibacterial traditional Chinese medicine(TCM)compounds.The objection of our research is to reshape the microenvironment in vivo environment to effectively screen out multi-target antibacterial Chinese herbal compound products.The method is to establish a flexible hydrogel drug sieve based on the bacterial cell interaction model and the micro-pattern imprint was re-engraved on the substrate to form a hydrogel cell culture chip with high through put and multiple geometric models.The infective interaction model of bacteria infected cells was established,and then Chinese herbal compound was added for treatment.The changes in the number of cells and bacteria were detected visually by immune-fluorescence technology,and the pharmacodynamic evaluation of TCM compounds was realized visually and quantitatively.The antibacterial effects of herbal compounds were detected by using the hydrogel cell sieve chip.The results showed the Niujiao Dihuang decoction,Qingying decoction and Qingwen Baidu decoction had significant antibacterial effects,among which Qingwen Baidu decoction had the best antibacterial effect and little damage to cells.The cytotoxicity of Pulsatilla decoction was high and the antibacterial activity was not significant.

关 键 词：水凝胶基底 抗菌中药复方 细胞筛药方法 可视化定量化处理 

分 类 号：S854.5[农业科学—临床兽医学]