基于药物浓度-免疫平衡-耐药性探讨芪归银方增敏左氧氟沙星的作用机制

The infection of drug-resistant pseudomonas aeruginosa (PAE) has become increasingly serious, and has been a major issue urgently to be solved in the international medical community. Studies have shown that immune imbalance is the key pathway for PAE infection. Chinese medicinal herbs combined with antibiotics can provide effective clinical strategies, but the mechanism is not clear. The preliminary study of this subject found that Qiguiyin prescription based on Fuzheng Touxie rule can regulate immune balance and sensitize levofloxacin for drug-resistant pseudomonas aeruginosa infection. To reveals the intrinsic mechanism that Qiguiyin prescription regulate immune balance and sensitize to antibiotics, the subject intends to use antibody microarray technology to study the occurrence and development of (resistant) PAE infection and the dynamic changes of inflammatory cytokines in rats after the interventions of different drugs, and construct the difference spectrum of immune factors. In addition, the subject intends to research the pharmacokinetic parameters of Qiguiyin-levofloxacin and levofloxacin alone, and analyze its correlation with drug resistance, and construct PK/PD model based on drug concentration-immune balance-drug resistance to predict the PK/PD parameters that prevent drug resistance. The internal mechanism of antibiotic sensitized by Qiguiyin prescription avoids PAE resistance was explained from the aspects of immune regulation and drug resistance prediction. The research of this subject can provide a scientific basis for the clinical application of traditional Chinese medicine in drug-resistant PAE infection, and provide effective strategies to solve the problem of drug-resistant infections.

耐药铜绿假单胞菌（PAE）感染日趋严峻，已成为国际医学界亟待解决的重大课题。研究表明免疫失衡是PAE感染发生的关键途径，中药联合抗生素可为其临床治疗提供有效策略，但作用机制不明确。本课题前期研究发现基于扶正透邪治则的芪归银方对耐药PAE感染可调控免疫平衡并增敏抗生素。因此，本课题拟采用抗体芯片技术研究（耐药）PAE感染发生、发展过程，以及不同药物干预大鼠体内炎性细胞因子水平的动态变化，构建免疫因子差异谱，揭示芪归银方调控免疫平衡增敏抗生素的内在机制；研究芪归银方-左氧氟沙星联用与左氧氟沙星单用的药代动力学参数，并分析其与耐药的相关性；构建基于药物浓度-免疫平衡-耐药性的多参数PK/PD结合模型，预测防止耐药发生的PK/PD参数，从免疫调控和耐药预测两个方面诠释芪归银方增敏抗生素防治PAE耐药的内在机制；为中医药治疗PAE耐药感染的临床应用提供科学依据，为解决细菌耐药感染问题提供有效策略。

铜绿假单胞菌（PA）耐药日趋严峻，已成为国际医学界亟待解决的重大课题，研究表明免疫失衡是其感染发生的关键，中药联合抗生素可为其临床治疗提供有效策略，但作用机制不明确。. 本课题以经口气管插管的方法建立了PA致大鼠肺炎模型；采用抗体芯片技术探究芪归银方联用左氧氟沙星对感染大鼠免疫指标的调控作用，比对不同给药方案的调控差异；采用UPLC-MS/MS技术比较左氧氟沙星单用及联用芪归银方的药代动力学差异，结合药动学建立了PK/PD模型评价联用芪归银方对左氧氟沙星抗PA的有效性，揭示药物浓度-免疫平衡与耐药性的相关性。结果表明：与左氧氟沙星单用相比，联合芪归银方能够于PA感染早期适度增加促炎细胞因子IL-1β、IL-6、TNF-α及IFN-γ的水平，后期能使促炎细胞因子的释放迅速下降，同时调控抗炎细胞因子IL-10呈现与促炎细胞因子相同的时相变化，控制促炎反应在“适度”水平，有益于调控机体炎症免疫反应的紊乱；药代动力学研究发现，与单用组相比，联用芪归银后不同致病菌感染大鼠体内的PK参数AUC和Cmax均降低，MRT(0-t)和Tmax均延长；药效学研究表明，联用组血清对耐药/敏感PA的抑菌率都显著高于单用组，与单用组相比，联用芪归银后大鼠血清中各种炎症因子的水平更加接近正常大鼠；联用组PA的生物膜形成量、生长曲线、毒力因子形成量均低于单用组，联用芪归银方能明显增强左氧氟沙星对PA生物膜的抑制作用。. 综上所述，耐药/敏感PA感染后会导致机体免疫失衡，联用芪归银方有助于调节促炎/抗炎细胞因子之间的动态平衡，增强左氧氟沙星防治耐药/敏感PA的作用。芪归银方能够改变左氧氟沙星在肺炎大鼠体内的药物浓度及达峰时间、清除率等药代参数；建立的PK/PD模型进一步证明了联用芪归银方的有效性。联用芪归银对PA的抑菌效果均优于单用，联用芪归银可以使左氧氟沙星对PA的MPC降低、MSW缩窄，从而延缓耐药突变PA菌株的产生。研究结果可为中药增敏抗生素作用机制阐释和临床给药方案的制定提供科学依据，为铜绿假单胞菌耐药感染的问题提供有效策略。