内化菌耐受性Tr值评估方法研究

doi:10.16431/j.cnki.1671-7236.2024.05.045

摘要/Abstract

摘要： 【目的】建立一种全面考虑宿主细胞、内化菌、抗菌药物三者互作的评估方法来精准比较内化菌介导的抗菌药物耐受情况。【方法】以IEC-6、RIMVECs和RAW264.7细胞为宿主细胞,以金黄色葡萄球菌和沙门菌作为内化菌,以氨苄西林、环丙沙星、四环素等7种化学药品及槲皮素和山柰酚等4种中药标准品为抗菌药物,检测抗菌药物细胞外最小杀菌浓度(MBC_EX),建立细菌-细胞感染互作模型,利用细菌菌落计数检测抗菌药物细胞内最小杀菌浓度(MBC_IN)。测定内化菌感染细胞的百分比(N)及药物治疗后内化菌感染细胞的百分比(N_D)。通过计算得到内化菌耐受值(Tr),对内化菌的抗菌药物耐受性进行评估。【结果】金黄色葡萄球菌对氨苄西林、环丙沙星、红霉素、多黏菌素、利福平、四环素、万古霉素7种化学药品和中药标准品黄芩苷均产生了耐受性(Tr值＞1),其中,对环丙沙星耐受性Tr值最高,耐受程度最严重;对槲皮素、山奈酚和儿茶素不产生耐受性(Tr值≤1)。不同内化菌对四环素的耐受性结果显示,金黄色葡萄球菌对四环素产生的耐受性Tr值大于沙门菌。不同宿主细胞对内化菌介导的抗菌药物耐受性的影响显示,IEC-6细胞中金黄色葡萄球菌对四环素的耐受性最高。【结论】耐受性Tr值评估方法不仅可有效评估内化菌对不同抗菌药物的耐受程度,还可比较不同宿主细胞对内化菌介导的抗菌药物耐受性的影响,将内化菌对抗菌药物产生的耐受性可视化和数据化地呈现出来,为临床精准用药提供指导。

关键词: 内化菌; 抗菌药物; 耐受性; 耐受值评估方法

Abstract: 【Objective】 This study was aimed to develope a comprehensive evaluation method that considered the interaction of host cells,internalized bacteria and antimicrobial agents to accurately compare internalized bacteria-mediated antimicrobial resistance.【Method】 Using IEC-6,RIMVECs and RAW264.7 cells as host cells,Staphylococcus aureus and Salmonella as internalized bacteria,seven chemical drugs including ampicillin,ciprofloxacin,tetracycline,and four traditional Chinese medicine standards including quercetin and kaempferol as antibacterial drugs,the minimum extracellular bactericidal concentration (MBC_EX) of antibiotics was detected,and the bacteria-cell interaction model was established.The minimum intracellular bactericidal concentration (MBC_IN) of antibiotics was detected by bacterial colony count.The percentage of endobacteria-infected cells (N) and the percentage of endobacteria-infected cells (N_D) after drug treatment were measured.The drug tolerance value (Tr) was calculated to evaluate the antimicrobial resistance mediated by internalized bacteria.【Result】 Staphylococcus aureus was resistant to ampicillin,ciprofloxacin,erythromycin,polymyxin,rifampin,tetracycline,vancomycin and baicalin (Tr＞1).Among them,the Tr value of ciprofloxacin was the highest and the tolerance was the most serious.It did not produce tolerance to quercetin,kaempferol and catechin (Tr≤1).The results of tolerance of different internalized bacteria to tetracycline showed that the Tr value of resistance of Staphylococcus aureus to tetracycline was greater than that of Salmonella.The influence of different host cells on internalized bacteria-mediated antimicrobial resistance showed that Staphylococcus aureus had the highest tolerance to tetracycline in IEC-6 cells.【Conclusion】 The Tr value evaluation method of tolerance could not only effectively evaluate the tolerance degree of internalized bacteria to different antibiotics,but also compare the influence of different host cells on the resistance to antibiotics mediated by internalized bacteria,and visually and data-present the tolerance of internalized bacteria to antibiotics,providing guidance for clinical precision medicine.

Key words: internalized bacteria; antibacterial drugs; tolerance; assessment of tolerance value

中图分类号:

孙志刚, 王潇, 司晓慧, 吕若一, 宋连宇, 刘晓晔. 内化菌耐受性Tr值评估方法研究[J]. 中国畜牧兽医, 2024, 51(5): 2245-2252.

SUN Zhigang, WANG Xiao, SI Xiaohui, LYU Ruoyi, SONG Lianyu, LIU Xiaoye. Study on Assessment of Internalized Bacteria-mediated Antimicrobial Tolerance Tr Value[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 2245-2252.

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