恩诺沙星联合用药对MRC-5细胞生长抑制的研究  

Evaluation of Joint Toxicity of Enrofloxacin and Three Antimicrobials on MRC-5 Cells

作　　者：栾业辉 程林丽[1] 赵军杰 丁宇丽 王文浩 黄家模 LUAN Yehui;CHENG Linli;ZHAO Junjie;DING Yuli;WANG Wenhao;HUANG Jiamo(College of Veterinary Medicine,China Agricultural University,Beijing 100193,China)

机构地区：[1]中国农业大学动物医学院,北京100193

出　　处：《中国畜牧兽医》2020年第11期3722-3730,共9页China Animal Husbandry & Veterinary Medicine

基　　金：国家重点研发计划(2018YFC1603005)。

摘　　要：本研究旨在评价恩诺沙星分别与3种抗菌药(环丙沙星、氟苯尼考和磺胺二甲嘧啶)的联合毒性。选用MRC-5细胞作为细胞模型模拟抗菌药的混合污染对肺细胞的损害,设置多个浓度梯度和混合比例,采用CCK-8方法检测4种抗菌药单药对细胞生长的抑制率及恩诺沙星分别与3种抗菌药混合后对细胞生长的抑制率,而后使用Chou-Talalay方法拟合中值效应曲线,计算出联合指数(combination index,CI)。结果显示,在检测浓度范围内,4种单药对MCR-5细胞的生长抑制率随药物浓度增加均呈阶梯状上升,其中氟苯尼考对MCR-5细胞的生长抑制率较低(<4.5%)。3种二元药物组合的混合毒性均表现出了浓度依赖性和混合比例的依赖性,联合使用恩诺沙星与环丙沙星在高、中剂量组下对MRC-5细胞表现协同毒性(CI<1),极低剂量组下表现为抑制毒性(CI>1);联合使用恩诺沙星与氟苯尼考对MRC-5细胞的毒性主要是相互抑制的(CI>1);联合使用恩诺沙星与磺胺二甲嘧啶对MRC-5细胞的毒性可能表现相互促进(CI<1),也可能表现相互抑制(CI>1),具体表现与给药剂量和浓度比值有关。本研究表明,在抗菌药的毒性评价中,评估抗菌药的联合毒性是有必要的,使用Chou-Talalay方法可快速高效地完成细胞水平上对抗菌药联合毒性的评估。The aim of this study was to evaluate the combined toxicity of enrofloxacin with each of three antimicrobials(ciprofloxacin,florfenicol and sulfamethazine).MRC-5 cells were used as a cell model to simulate the damage of lung cells caused by mixed contamination of antimicrobials.Multiple concentration gradients and mixing ratios were set.CCK-8 method was used to determine the inhibition rate of cell growth caused by four antimicrobials and the inhibition rate of cell growth caused by enofloxacin mixed with three antimicrobials,respectively.Then Chou-Talalay method was used to fit the median effect plot and to calculate the combination index(CI)value.The results showed that the growth inhibition rates of MCR-5 cells caused by four single drugs went up in a step-like manner with the increase of drug concentration in the tested concentration range,among them,the growth inhibitory rate of MCR-5 cells by florfenicol was low(<4.5%).The combined toxicity of the three binary combinations showed a concentration-dependent and mixing-ratio dependence.Mixing enrofloxacin and ciprofloxacin showed synergistic toxicity(CI<1)on MRC-5 cells at high and middle concentration groups,and antagonistic toxicity(CI>1)at the very-low-concentration groups.Mixing ciprofloxacin and florfenicol mainly showed an inhibited toxicity(CI>1).Binary combination enrofloxacin and sulfamethazine might present either a synergistic joint toxicity(CI<1)or an antagonistic joint toxicity(CI>1)as the concentration and mixing ratio changing.This study showed that it was necessary to assess the combined toxicity of antimicrobials in the toxicity evaluation of antimicrobials.Using Chou-Talalay method,the joint toxicity of multiple antimicrobials could be quickly and efficiently determined at cellular level.

关 键 词：恩诺沙星 联合毒性 Chou-Talalay方法 

分 类 号：S859.79[农业科学—临床兽医学]