[1]周桓宇,伍慧妍,关艺琳.中药苦参协同增强对替加环素耐药CRKP的体外抑菌作用及机制研究[J].现代检验医学杂志,2025,40(06):176-180.[doi:10.3969/j.issn.1671-7414.2025.06.032]
　ZHOU Huanyu,WU Huiyan,GUAN Yilin.Research on the Synergistic Enhancement of the In Vitro Bactericidal Effect of Tigecycline-resistant CRKP by Traditional Chinese Medicine Radix Sophorae Flavescentis and Its Mechanism[J].Journal of Modern Laboratory Medicine,2025,40(06):176-180.[doi:10.3969/j.issn.1671-7414.2025.06.032]
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中药苦参协同增强对替加环素耐药CRKP的体外抑菌作用及机制研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年06期
页码:
176-180
栏目:
论著
出版日期:
2025-11-15

文章信息/Info

Title:
Research on the Synergistic Enhancement of the In Vitro Bactericidal Effect of Tigecycline-resistant CRKP by Traditional Chinese Medicine Radix Sophorae Flavescentis and Its Mechanism
文章编号:
1671-7414（2025）06-176-05
作者:
周桓宇1,2，伍慧妍1,2，关艺琳1,2
1.广州医科大学附属第五医院，广州510700;2.广州医科大学金域检验学院，广州 510180
Author(s):
ZHOU Huanyu1,2，WU Huiyan1,2，GUAN Yilin1,2
1.the Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou 510700, China;2.KingMed School of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510180, China
关键词:
苦参; 替加环素; 耐碳青霉烯肺炎克雷伯菌; AcrAB-Tolc外排泵
分类号:
R285；R446.5
DOI:
10.3969/j.issn.1671-7414.2025.06.032
文献标志码:
A
摘要:
目的探究苦参对替加环素耐药的耐碳青霉烯类肺炎克雷伯菌（CRKP）的体外抑菌作用，阐明苦参如何通过外排泵途径恢复CRKP对替加环素的敏感度。方法采用微量肉汤稀释法体外诱导CRKP标准菌株BAA1705对替加环素耐药建立模型，并通过外排泵抑制剂验证其外排泵表达情况；微量棋盘稀释法测定两药联用的最低抑菌浓度（MIC）并计算部分抑菌浓度（FIC）指数；荧光实时定量PCR（qRT-PCR）分析苦参处理前后外排泵基因AcrB的表达量；罗丹明6G外排试验验证外排泵活性。结果人工造模T-BAA1705外排泵抑制试验结果显示其外排泵过度表达。联合抑菌试验结果计算FIC值为0.3752，提示苦参与替加环素联合使用具有协同作用。qRT-PCR结果显示，苦参单药组AcrB水平低于无药物处理组（0.018±0.002vs0.040±0.001），联合用药组的AcrB水平（0.009±0.001），RamA水平（0.184±0.003）以及MarA水平（0.013±0.001）均低于无药物处理组（0.040±0.001，0.387±0.016，0.125±0.007），而AcrR水平高于无药物处理组（0.388±0.001vs0.288±0.001），差异具有统计学意义（t=12.28～47.12，均P＜0.05）；苦参的单药组外排泵活性和联合用药组外排泵活性均低于未经过药物处理的T-BAA1705（60.667±0.448，60.267±0.376vs65.133±0.296），差异具有统计学意义（t=8.31，10.17，均P＜0.05）。结论?苦参能恢复耐替加环素的CRKP对替加环素的敏感性，同时与替加环素联合使用能有效抑制细菌的生长。其中的机制可能是苦参抑制AcrAB-TolC外排泵的表达。
Abstract:
Objective To investigate the in vitro antibacterial effect of Radix Sophorae Flavescentis on tigecycline-resistant carbapenem-resistant Klebsiella pneumonia(CRKP) and elucidate how Radix Sophorae Flavescentis restores the sensitivity of CRKP to tigecycline through the efflux pump pathway. Methods The CRKP standard strain BAA1705 was induced to be tigecycline-resistant using the microdilution broth method, and the expression of efflux pumps was verified by efflux pump inhibitors. The fractional inhibitory concentration index (FIC) was calculated using the microdilution checkerboard method to determine the minimum inhibitory concertration(MIC) of the two drugs in combination. Fluorescent real-time quantitative PCR (qRT-PCR) was used to analyze the expression of the AcrB gene before and after Radix Sophorae Flavescentis treatment, and the e?ux pump activity was validated by the Rhodamine 6G e?ux assay. Results The e?ux pump inhibition experiment on the artificially induced T-BAA1705 strain showed overexpression of e?ux pumps. The combined antibacterial test results indicated synergy between Radix Sophorae Flavescentis and tigecycline (FIC=0.375 2 ). The qRT-PCR results showed that the AcrB levels in the Radix Sophorae Flavescentis monotherapy group were lower than in the untreated group(0.018±0.002 vs 0.040±0.001), and the AcrB levels(0.009±0.001), RamA levels(0.184±0.003) and MarA(0.013±0.001) levels in the combination therapy group were all lower than in the untreated group(0.040±0.001, 0.387±0.016, 0.125±0.007), while the AcrR level was higher than in the untreated group(0.388±0.001 vs 0.288±0.001), and the differences were statistical significance (t=12.28 ~ 47.12, all P<0.05); the e?ux pump activity in the Radix Sophorae Flavescentis monotherapy group and the combination therapy group was lower than in the untreated T-BAA1705(60.667±0.448, 60.267±0.376, 65.133±0.296), and the differences were statistical significance (t=8.31,10.17, all P< 0.05). Conclusion Radix Sophorae Flavescentis can restore the sensitivity of tigecycline-resistant CRKP to tigecycline and effectively inhibit bacterial growth when used in combination with tigecycline. The mechanism may involve Radix Sophorae Flavescentis inhibiting the expression of the AcrAB-TolC e?ux pump.

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备注/Memo

备注/Memo:
基金项目：广东省医学科学技术研究基金项目(B2021440)；广州市中医药和中西医结合科技项日(202324011017）。
作者简介：周桓宇（2001-），男，在读研究生，研究方向：临床检验诊断学，E-mail：17322833487@163.com。
通讯作者：关艺琳（1992-），女，本科，检验技师，研究方向：细菌耐药和异质性耐药机制，E-mail：colin.kwan@163.com。
更新日期/Last Update: 2025-11-15