芪归银方对耐亚胺培南铜绿假单胞菌耐药作用的影响

目的观察芪归银方体外逆转耐亚胺培南铜绿假单胞菌耐药的作用。方法采用血清药理学方法制备芪归银方含药血清;常规肉汤培养耐亚胺培南铜绿假单胞菌(1643菌株),分为含药血清组(M-H肉汤培养基0.8ml+含药血清0.2ml)、空白血清组(M-H肉汤培养基0.8ml+空白血清0.2ml)、阳性对照组(M-H肉汤培养基1ml),各组分别传代干预至第8代,用二倍稀释法测定细菌对亚胺培南的最小抑菌浓度(MIC),并比较抑菌环直径;耐亚胺培南铜绿假单胞菌冻融离心,收集酶粗提液,以头孢噻吩为底物,分为含药血清组(0.1mmol/L头孢噻吩95μl+3μl含药血清+2μl酶粗提液)和空白血清组(0.1mmol/L头孢噻吩95μl+3μl空白血清+2μl酶粗提液),测定耐药铜绿假单胞菌β-内酰胺酶水解速率。结果含药血清组耐亚胺培南铜绿假单胞菌对亚胺培南的MIC为8μg/ml,空白血清组与阳性对照组均为16μg/ml;含药血清组抑菌环直径显著大于阳性对照组(P<0.05);含药血清组β-内酰胺酶水解速率显著低于空白血清组(P<0.05)。结论在芪归银方含药血清作用下,耐亚胺培南铜绿假单胞菌对亚胺培南的敏感性一定程度上得到了恢复,并且可以减慢β-内酰胺酶的水解速率,提示抑制β-内酰胺酶水解可能是芪归银方干预耐亚胺培南铜绿假单胞菌耐药的机制之一。

Effect of Qiguiyin Formula on the Imipenem-Resisting Pseudomonas Aeruginosa

Objective To observe the effect of Qiguiyin Formula Formula of Huangqi(Radix Astrangali),Danggui(Angelica sinesis) and Jinyinhua(Lonicera Japonica)] on imipenem-resisting pseudomonas aeruginosa.Methods Serum pharmacological method was used to prepare drug-containing serum of Qiguiyin Formula.Imipenem-resisting pseudomonas aeruginosa(1643 strains) was cultured with conventional broth and was divided into the drug-containing group(0.8ml of MH broth+0.2ml of drug-containing serum),blank ser um group(0.8ml of MH broth+0.2ml of blank serum) and the positive control group (1ml of MH broth).Three groups were given passaged intervention to the 8th gener ation.The minimal inhibitory concentration(MIC) value of strains in each group to imipenem was determined with double dilution method and the inhibition zone diameter was compared.Imipenem-resisting pseudomonas aeruginosa strains was frozen,thawed and centrifuged.The enzyme crude extracts was collected.Cephaloridine was used as substr ate and divided into the drug-containing serum group(95μl of 0.1mM/L cephaloridine+3μ l of drug-containing serum+2μl of enzyme crude extracts) and blank serum group(95μl o f 0.1mM/L cephaloridine+3μl of blank serum+2μl of enzyme crude extracts).The hyd r olysis rate of β-lactamase was determined.Results Th e MIC value of imipenem-resisting pseudomonas aeruginosa to imipenem in the drug-containing serum group,blank serum group an d positive control group was 8μg/ml,16μg/ml and 16μg/ml respectively.The inhibition zone diameter in the drug-containing serum group was significantly larger than that in the positive control g roup(P<0.05).The hydrolysis rate of β-lactamase in the drug-containing serum group was significantly lower than that in the blank serum group(P<0.05).Conclusion The sensitivity of imipenem-resisting pseudomonas aeruginosa to imipenem could be restored by the drug-containing serum of Qiguiyin Formula.The Qiguiyin Formula-containing serum of could reduce the hydrolysis rate of β-lactamase,which maybe the mechanism of its reversal effect on the resisting of imipenem-resistant ps eudomonas aeruginosa.