胃肠道解痉药匹维溴铵对表皮葡萄球菌的体外和体内抗菌活性研究  

In vitro and in vivo antimicrobial activity of gastrointestinal antispasmodic drug pinaverium bromide against Staphylococcus epidermidis

作　　者：彭程[1,2] 彭聪 侯正利[1] PENG Cheng;PENG Cong;HOU Zheng-li(Department of Laboratory Medicine,Changsha Hospital of Hunan Normal University,Changsha 410006,China;Department of Laboratory Medicine,The Fourth Hospital of Changsha,Changsha 410006,China)

机构地区：[1]湖南师范大学附属长沙医院检验科,湖南长沙410006 [2]长沙市第四医院检验科,湖南长沙410006

出　　处：《中国感染控制杂志》2024年第9期1077-1083,共7页Chinese Journal of Infection Control

基　　金：湖南省自然科学基金项目(2023JJ30060)。

摘　　要：目的研究匹维溴铵(PVB)对表皮葡萄球菌(表葡)的体外和体内抗菌活性。方法收集长沙市某医院2022年1-12月住院患者血液分离的表葡。采用微量肉汤稀释试验和纸片扩散法检测表葡对PVB的敏感性,通过时间-杀菌曲线检测PVB抗菌效果的时间和浓度依赖性,透射电镜观察PVB处理后细菌的超微结构改变,结晶紫染色试验检测PVB对表葡生物膜的抑制和清除效果,采用棋盘稀释法研究PVB与抗菌药物的联合抗菌效果。构建皮肤脓肿感染模型,检测PVB的体内抗菌活性。结果药敏试验结果显示,PVB对表葡标准菌株RP62A、ATCC 12228的最低抑菌浓度(MIC)、最低杀菌浓度(MBC)均分别为8、16μg/mL;对表葡临床菌株的MIC、MBC分别为4~8μg/mL、8~16μg/mL。纸片扩散法结果显示,与未加药的对照组(0.60±0)cm相比,0.2 mg PVB出现明显抑菌圈[(2.26±0.09)cm;t=45.34,P<0.001],且抑菌圈直径随着PVB药量增加而增大。时间-杀菌曲线结果提示,PVB具有杀菌活性,且随着药物浓度和作用时间的增加而增强。透射电镜观察发现PVB可明显破坏表葡的正常结构,导致细菌水肿和裂解。此外,1×MIC的PVB还可显著抑制表葡生物膜的形成,使其生物膜的形成量(A 570 nm)从(2.30±0.18)减少到(0.47±0.11;t=14.85,P<0.001)。同时,1×MIC的PVB还可有效破坏已形成的生物膜,使生物膜的量从(2.64±0.10)减少到(1.77±0.30;t=4.76,P=0.009)。PVB与阿米卡星和庆大霉素联用具有协同抗菌活性,其协同抑菌指数分别为0.50、0.31。动物模型发现10 mg/kg体重的PVB可使脓肿面积从(68.83±10.68)mm 2减少到(35.50±10.58)mm 2(t=6.52,P<0.001),且使脓肿中的活菌量从(6.11±0.55)lg(CFU/脓肿)减少到(3.60±0.34)lg(CFU/脓肿)(t=3.08,P=0.014)。苏木精-伊红染色发现PVB用药组皮肤脓肿中的炎性细胞浸润相比对照组明显减少,趋于正常。结论PVB对表葡具有明显的体外和体内抗菌活性,有望成为表葡相关感染的替代治疗途�Objective To explore the in vitro and in vivo antimicrobial activity of pinaverium bromide(PVB)against Staphylococcus epidermidis(S.epidermidis).Methods S.epidermidis isolated from blood specimens of hospitalized patients in a hospital in Changsha from January to December 2022 were collected.Susceptibility test of S.epidermidis to PVB was performed using broth microdilution method and disc diffusion method.The time-and concentration-dependent antimicrobial activity of PVB were determined by time-killing assay.Ultrastructural changes in bacteria after PVB treatment was observed by transmission electron microscope.The inhibitory and clearance effects of PVB on S.epidermidis biofilm were detected by crystal violet staining test.The combined antimicrobial effect of PVB and antimicrobial agents was studied through microdilution checkerboard technique.A skin abscess infection model was constructed to detect the in vivo antimicrobial activity of PVB.Results Antimicrobial susceptibility testing results showed that the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of PVB against the standard strains RP62A and ATCC 12228 were 8 and 16μg/mL,respectively;The MIC and MBC of clinical strains of S.epidermidis were 4-8μg/mL and 8-16μg/mL,respectively.Disc diffusion method results showed that compared with the untreated control group(0.60±0)cm,0.2 mg PVB treatment showed a significant inhibitory zone([2.26±0.09]cm;t=45.34,P<0.001),and the diameter of inhibitory zone increased with the increase of PVB dosage.The time-killing curves indicated PVB had bactericidal activity,which enhanced with increased concentration and action duration.Transmission electron microscope observed that PVB could significantly damage the normal structure of S.epidermidis,leading to bacterial edema and lysis.In addition,at the concentration of 1×MIC,PVB could significantly inhibit the formation of S.epidermidis biofilm,reducing the amount of biofilm formation(A 570 nm)from(2.30±0.18)to(0.47±0.11;t=14.85,P<0.001).M

关 键 词：匹维溴铵 药物重利用 表皮葡萄球菌 生物膜 脓肿感染模型 抗菌活性 

分 类 号：R378.11[医药卫生—病原生物学]