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目的研究加味银翘散对铜绿假单胞菌外膜蛋白OprD2的干预作用。方法分别用加味银翘散原液、亚胺培南和加味银翘散原液+亚胺培南作用于铜绿假单胞菌,检测各组药物作用前后最低抑菌浓度值对亚胺培南耐药性的变化;利用荧光定量逆转录PCR检测外膜蛋白OprD2基因表达量的变化。结果 7株铜绿假单胞菌经加味银翘散原液+亚胺培南联合作用后,亚胺培南最低抑菌浓度值比药物单纯作用时均降低,下降幅度最大的达16倍;除PAE4、PAB5、PAC6三株菌株经药物作用后OprD2基因表达量变化均增加,差异均有统计学意义(P<0.05);PAA2、PAA9、PAE7、PAH3四株菌OprD2基因表达量变化均为0,差异均无统计学意义。结论亚胺培南和加味银翘散联合作用能降低铜绿假单胞菌对亚胺培南的耐药性;PAA2、PAA9、PAE7、PAH3四株菌对亚胺培南耐药不是由OprD2数量减引少起的,可能存在其他的耐药机制;PAE4、PAB5和PAC6经药作物用后,亚胺培南最低抑菌浓度值降低,外膜通透性均增加,说明该三株菌对亚胺培南耐药是由OprD2数量减少引起的。
Objective To study the effect of Jiawei Yinqiaosan on the outer membrane protein OprD2 of Pseudomonas aeruginosa. Methods Pseudomonas aeruginosa was treated with Jiawei Yinqiaosan stock solution, Imipenem and Jiawei Yinqiao powder stock solution and Imipenem, respectively. The MIC was tested for imipenem resistance before and after the drug treatment. The change of sex; the use of fluorescent quantitative reverse transcription PCR detection of the outer membrane protein OprD2 gene expression changes. Results After 7 strains of Pseudomonas aeruginosa were combined with Jiawei Yinqiao bulk solution and imipenem, the MIC value of imipenem was lower than that of the drug alone, and the greatest decrease was 16 times; except for PAE4. The expression of OprD2 gene in the three strains of PAB5 and PAC6 was increased after drug treatment, and the difference was statistically significant (P<0.05). The expression levels of OprD2 gene in PAA2, PAA9, PAE7 and PAH3 strains were all 0. The difference was not statistically significant. Conclusion The combined effect of imipenem and Jiawei Yinqiaosan can reduce the resistance of P. aeruginosa to imipenem; PAA2, PAA9, PAE7 and PAH3 are resistant to imipenem by OprD2. If there are fewer subtractions, there may be other mechanisms of resistance; PAE4, PAB5, and PAC6 are reduced by the minimum inhibitory concentration of imipenem and the permeability of the outer membrane after the use of the crops, indicating that the three strains Imipenem resistance is caused by a decrease in the number of OprD2.