PDF(Pubmed)
Abstract：
Pseudomonas aeruginosa (P. aeruginosa) is a common pathogen associated with biofilm infections, which can lead to persistent infections. Therefore, there is an urgent need to develop new anti-biofilm drugs. DZ2002 is a reversible inhibitor that targets S-adenosylhomocysteine hydrolase and possesses anti-inflammatory and immune-regulatory activities. However, its anti-biofilm activity has not been reported yet.
Therefore, we investigated the effect of DZ2002 on P. aeruginosa PAO1 biofilm formation by crystal violet staining (CV), real-time quantitative polymerase chain reaction (RT-qPCR) and confocal laser scanning microscopy (CLSM). The results indicated that although DZ2002 didn\'t affect the growth of planktonic PAO1, it could significantly inhibit the formation of mature biofilms. During the inhibition of biofilm formation by DZ2002, there was a parallel decrease in the synthesis of alginate and the expression level of alginate genes, along with a weakening of swarming motility. However, these results were unrelated to the expression of lasI, lasR, rhII, rhIR. Additionally, we also found that after treatment with DZ2002, the biofilms and extracellular DNA content of PAO1 were significantly reduced. Molecular docking results further confirmed that DZ2002 had a strong binding affinity with the active site of S-adenosylhomocysteine hydrolase (SahH) of PAO1.
In summary, our results indicated that DZ2002 may interact with SahH in PAO1, inhibiting the formation of mature biofilms by downregulating alginate synthesis, extracellular DNA production and swarming motility. These findings demonstrate the potential value of DZ2002 in treating biofilm infections associated with P. aeruginosa.
Therefore, we investigated the effect of DZ2002 on P. aeruginosa PAO1 biofilm formation by crystal violet staining (CV), real-time quantitative polymerase chain reaction (RT-qPCR) and confocal laser scanning microscopy (CLSM). The results indicated that although DZ2002 didn\'t affect the growth of planktonic PAO1, it could significantly inhibit the formation of mature biofilms. During the inhibition of biofilm formation by DZ2002, there was a parallel decrease in the synthesis of alginate and the expression level of alginate genes, along with a weakening of swarming motility. However, these results were unrelated to the expression of lasI, lasR, rhII, rhIR. Additionally, we also found that after treatment with DZ2002, the biofilms and extracellular DNA content of PAO1 were significantly reduced. Molecular docking results further confirmed that DZ2002 had a strong binding affinity with the active site of S-adenosylhomocysteine hydrolase (SahH) of PAO1.
In summary, our results indicated that DZ2002 may interact with SahH in PAO1, inhibiting the formation of mature biofilms by downregulating alginate synthesis, extracellular DNA production and swarming motility. These findings demonstrate the potential value of DZ2002 in treating biofilm infections associated with P. aeruginosa.
摘要：
■铜绿假单胞菌（P.铜绿假单胞菌）是与生物膜感染相关的常见病原体，会导致持续性感染.因此,迫切需要开发新的抗生物膜药物。DZ2002是靶向S-腺苷同型半胱氨酸水解酶的可逆抑制剂,具有抗炎和免疫调节活性。然而,其抗生物膜活性尚未报道。
■因此，我们通过结晶紫染色（CV）研究了DZ2002对铜绿假单胞菌PAO1生物膜形成的影响，实时定量聚合酶链反应（RT-qPCR）和共聚焦激光扫描显微镜（CLSM）。结果表明,DZ2002对浮游PAO1的生长无影响,但能显著抑制成熟生物膜的形成。在DZ2002抑制生物膜形成的过程中，藻酸盐的合成和藻酸盐基因的表达水平平行下降，伴随着蜂群运动的减弱。然而,这些结果与lasI的表达无关，激光,rhII,rhIR.此外,我们还发现，用DZ2002处理后，PAO1的生物膜和细胞外DNA含量显着降低。分子对接结果进一步证实DZ2002与PAO1的S-腺苷同型半胱氨酸水解酶(SahH)的活性位点具有强结合亲和力。
■总之，我们的结果表明DZ2002可能与PAO1中的SahH相互作用，通过下调藻酸盐的合成来抑制成熟生物膜的形成，细胞外DNA产生和成群运动。这些发现证明了DZ2002在治疗与铜绿假单胞菌相关的生物膜感染中的潜在价值。
■因此，我们通过结晶紫染色（CV）研究了DZ2002对铜绿假单胞菌PAO1生物膜形成的影响，实时定量聚合酶链反应（RT-qPCR）和共聚焦激光扫描显微镜（CLSM）。结果表明,DZ2002对浮游PAO1的生长无影响,但能显著抑制成熟生物膜的形成。在DZ2002抑制生物膜形成的过程中，藻酸盐的合成和藻酸盐基因的表达水平平行下降，伴随着蜂群运动的减弱。然而,这些结果与lasI的表达无关，激光,rhII,rhIR.此外,我们还发现，用DZ2002处理后，PAO1的生物膜和细胞外DNA含量显着降低。分子对接结果进一步证实DZ2002与PAO1的S-腺苷同型半胱氨酸水解酶(SahH)的活性位点具有强结合亲和力。
■总之，我们的结果表明DZ2002可能与PAO1中的SahH相互作用，通过下调藻酸盐的合成来抑制成熟生物膜的形成，细胞外DNA产生和成群运动。这些发现证明了DZ2002在治疗与铜绿假单胞菌相关的生物膜感染中的潜在价值。
