目的 建立一种AmpC酶抑制剂的快速筛选方法，为治疗由耐药菌传播引起的感染提供筛选依据。方法 以阿维巴坦钠为阳性对照，采用紫外分光光度法检测反应体系的吸光度变化，通过6种不同底物的筛选比较及反应条件的优化，并对136种中药提取物进行筛选评价。结果 最佳体系为波长489 nm、底物质量浓度0.1 mg/mL、酶浓度400 U/mL、温度30℃、反应时间300 s。136种中药水提物中有12种中药水提物对AmpC酶的抑制率大于70%，其中桑寄生、赤芍、红景天、千斤拔、石见穿5种中药水提物的抑制率大于80%；添加桑寄生可明显提高青霉素的协同抗菌效应，最低抑菌浓度（minimal inhibit concentration，MIC）由2 mg/mL降低至0.8 mg/mL。结论 建立的AmpC酶抑制剂筛选方法快速、高效、可信度高，中药是抗耐药菌感染治疗的重要资源。

Objective To establish a rapid screening method for AmpC enzyme inhibitors to provide a basis for screening drugs in treatment of infections caused by spread of drug-resistant bacteria. Methods Avibactam sodium was used as a positive control, and the absorbance changes of reaction system were detected by UV spectrophotometry. Six different substrates were compared for screening, and the reaction conditions were optimized. A total of 136 Chinese medicinal materials extracts were screened and evaluated. Results The optimal system was wavelength 489 nm, substrate concentration 0.1 mg/mL, enzyme concentration 400 U/mL, temperature 30℃, and reaction time 300 s. Among the 136 aqueous extracts of Chinese medicinal materials, 12 extracts showed AmpC enzyme inhibitory activities greater than 70%, and five extracts including Taxillus chinensis, Paeonia veitchii, Rhodiola crenulata, Flemingia philippinensis and Salvia chinensis had inhibitory activities greater than 80%, and the addition of T. chinensis significantly improved the synergistic antibacterial effect of penicillin, minimal inhibit concentration (MIC) was changed from 2 mg/mL to 0.8 mg/mL. Conclusion The established screening method for AmpC enzyme inhibitors is rapid, efficient and highly reliable, and traditional Chinese medicines are important resources for the treatment of infections caused by drug-resistant bacterial.

R285.51

国家自然基金资助项目（32160021）；甘肃省重点研发计划项目（21YF5FA084，22YF7NA176）；温州市科技局项目（2022Y0883）