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目的:研究亚致死剂量的云南白药水提取物(Aquatic Extract of Yunnan Baiyao,AEYB)对铜绿假单胞菌(Pseudomonas Aeruginosa,PA)AlgR1-AlgR2双组份调控系统的磷酸化、海藻酸盐的生物合成及生物膜形成的影响。方法:通过放射性同位素标记方法研究AEYB对AlgR1-AlgR2磷酸化的影响;采用real-time PCR分析YBX海藻酸盐合成酶基因algD转录的影响;用咔唑法和结晶紫法分别对海藻酸盐和静态生物膜进行定量测定。结果:AEYB并不抑制AlgR2的自磷酸化,但能抑制AlgR2的磷酸基团向AlgR1转移,从而抑制AlgR1的磷酸化。AEYB可显著下调algD基因的转录,并抑制alginate的合成,最终减少PA静态生物膜的形成。结论:云南白药可通过抑制AlgR1-AlgR2系统的磷酸化的途径来减少PA的alginate的生物合成,对PA生物膜的形成具有抑制作用。
OBJECTIVE: To investigate the effect of sub-lethal doses of AEYB on the phosphorylation of AlgR1-AlgR2 two-component regulatory system of Pseudomonas aeruginosa (PA), alginate bioaccumulation Synthesis and biofilm formation. Methods: The effect of AEYB on the phosphorylation of AlgR1-AlgR2 was studied by radioisotope labeling. The transcription of algD gene was analyzed by real-time PCR. The effects of alginate and Static biofilm for quantitative determination. RESULTS: AEYB did not inhibit the autophosphorylation of AlgR2 but inhibited the phosphorylation of AlgR1 by inhibiting the transfer of AlgR2 to AlgR1. AEYB significantly down-regulated algD gene transcription and inhibited the synthesis of alginate, ultimately reducing the formation of PA static biofilms. CONCLUSION: Yunnan Baiyao can inhibit the biosynthesis of alginate by inhibiting the phosphorylation of AlgR1-AlgR2 system, which has an inhibitory effect on the formation of PA biofilm.