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目的研究鱼腥草素钠(SH)对铜绿假单胞菌(Pseudomonas aeruginosa,PA)群感效应系统的影响,以期探明SH抗PA的作用机制。方法微量稀释法测定SH和阿奇霉素(AZM)对PA的最小抑菌浓度(MIC);紫外分光光度法测定SH对PA LasA蛋白酶活性和绿脓菌素生成的影响;平板法观察PA的形态;半定量PCR法和实时荧光定量PCR(q RT-PCR)法检测群感效应相关的基因lasI、rhlI、lasR、rhlR、phzM、pqsA、pslA、lasA、las B、toxA表达量的变化。结果 SH和AZM对PA的MIC分别为512μg/m L和64μg/m L;紫外分光光度法结果显示SH能够抑制LasA蛋白酶的活性和绿脓菌素的合成;SH能够显著影响PA的形态;半定量PCR和q RT-PCR结果显示在SH作用下lasI、rhlI、lasR、lasA、pslA、phz M基因表达显著发生变化。结论 SH具有抗PA群感效应属性,通过抑制PA的群感效应系统发挥抗菌作用。
Objective To study the effect of sodium houttuyfonate (SH) on the flu-susceptibility of Pseudomonas aeruginosa (PA) in order to explore the mechanism of action of SH against PA. Methods The minimal inhibitory concentration (MIC) of SH and azithromycin (AZM) on PA was determined by microdilution method. The effect of SH on PA LasA protease activity and pyocyanin production was determined by ultraviolet spectrophotometry. The morphology of PA was observed by plate method. Quantitative PCR and real-time quantitative PCR (q RT-PCR) were used to detect the expression of lasI, rhlI, lasR, rhlR, phzM, pqsA, pslA, lasA, Results The MICs of SH and AZM against PA were 512 μg / m L and 64 μg / m L, respectively. The results of UV spectrophotometry showed that SH inhibited the activity of LasA protease and pyocyanin synthesis. SH could significantly affect the morphology of PA. Quantitative PCR and q RT-PCR results showed that the expression of lasI, rhlI, lasR, lasA, pslA and phz M genes under the action of SH significantly changed. Conclusion SH has the anti-PA group flu effect property, and exerts antibacterial effect by inhibiting the group flu effect of PA.