木犀草素抑制NorM外排泵逆转副溶血弧菌耐药性的分子机制研究

群体感应系统在细菌耐药性调控中发挥重要作用。前期研究显示木犀草素可通过抑制NorM外排泵逆转副溶血弧菌耐药性，该过程中发现LuxS/AI-2群体感应系统基因luxS的表达显著降低。推测木犀草素抑制副溶血弧菌LuxS/AI-2系统可能是抑制NorM外排泵的重要机制。本项目拟通过研究木犀草素作用下LuxS基因和蛋白的表达模式，确证木犀草素与LuxS的关系；通过比较luxS缺失前后细菌胞内抗生素蓄积量变化、luxS过量表达时norM缺失株和野生株对抗生素的敏感性差异，明确LuxS对NorM外排泵的调控作用；通过研究NorM在转录和翻译水平上对LuxS的分子响应及NorM与LuxS的蛋白互作关系，解析LuxS对NorM的调控机制。研究结果可阐明木犀草素抑制NorM外排泵的分子调控机制，可为采取合理的外排泵抑制策略提供依据，为耐药副溶血弧菌防控提供新思路。

Quorum sensing system plays an important role in the regulation of bacterial resistance. Previous studies have shown that luteolin can reverse the resistance of Vibrio parahaemolyticus by inhibiting the NorM efflux pump. During this process, it was found that the expression of LuxS/AI-2 quorum sensing system gene luxS was significantly reduced. It is speculated that luteolin inhibits the LuxS/AI-2 system of Vibrio parahaemolyticus may be an important mechanism to inhibit NorM efflux pump. This project intends to confirm the relationship between luteolin and LuxS by studying the expression patterns of LuxS at the transcription and translation levels under the action of luteolin. Then it will compare the accumulation of intracellular antibiotics before and after luxS deletion and the difference in drug sensitivity between norM-deficient strains and wild strains when luxS is overexpressed to clarify the regulating effect of LuxS on NorM efflux pump. Finally, it will study the molecular response of NorM to LuxS at the transcription and translation level and the protein interaction between NorM and LuxS to analyze the regulatory mechanism of LuxS on NorM. The research results can clarify the molecular regulation mechanism of luteolin inhibiting NorM efflux pump, provide a basis for adopting reasonable efflux pump inhibition strategies, and provide new ideas for the prevention and control of drug-resistant Vibrio parahaemolyticus.