基于β-内酰胺酶和外膜蛋白探讨扶正透邪方对铜绿假单胞菌耐药性的作用机制

A serious threat to human health of drug-resistant bacteria and caused heavy economic burden, has become a difficult problem to be solved at home and abroad.Because of the complex mechanism of bacterial resistance, is still a lack of effective drugs and methods.The subject of traditional Chinese medicine based on the advantages, to drug-resistant bacteria with Pseudomonas aeruginosa representative as the research object, the main mechanism of resistance as research tools, with characteristics of TCM prescription, on the basis of previous studies, from the Fuzheng evil delaying / mechanism and centralizer, reversal of Pseudomonas aeruginosa the effect of antibiotic resistance through evil component compatibility effect and in vivo intervention effect on bacterial drug resistance and other aspects of the in-depth study, research and development for the effective preparation of traditional Chinese medicine and clinical drug resistant bacteria infection therapy of traditional Chinese medicine and drug selection to provide a scientific basis, so as to improve the efficacy of traditional Chinese medicine in prevention and treatment of drug-resistant bacteria infection, fully display the characteristics and advantages of Chinese medicine traditional Chinese medicine industry, to promote the development of drug-resistant bacteria infection.

耐药菌感染对人类健康的严重威胁及其导致的沉重经济负担，已经成为国内外亟待解决的棘手难题。由于细菌产生耐药的机制复杂，目前仍缺乏有效的药物和措施。本课题基于中医药的自身优势，以耐药菌中具有代表性的铜绿假单胞菌为研究对象，以其主要耐药机制为研究手段，切合中医病机特点组方，在前期研究的基础上，从扶正透邪方延缓/逆转铜绿假单胞菌耐药性的作用机制及其扶正、透邪组分配伍作用和体内干预作用对细菌耐药性的影响等方面开展深入研究，为中药有效制剂的研发以及临床上抗耐药菌感染中医治法和药物选择提供科学依据，进而提高中医药防治耐药菌感染疗效，充分显示中医药的特色与优势，推动抗耐药菌感染中药产业化发展。

本课题按计划书已完成全部研究内容，并延伸开展了部分工作：采用iTRAQ 方法及传统β-内酰胺酶水解速率检测方法等，研究了芪归银方基于β-内酰胺酶和外膜蛋白对铜绿假单胞菌耐药性影响的作用机制。获得如下结果：①芪归银干预的各组鼠含药血清均能有效延缓泰能诱导的铜绿假单胞菌耐药，为临床亚胺培南的用药赢得时间提供实验依据；②以系列监测的β-内酰胺酶水解速率结果，证实各组芪归银含药血清均能有效降低耐药铜绿假单胞菌中β-内酰胺酶的水解速率，从而明确了芪归银对PA菌耐药基于β-内酰胺酶的影响机制；③综合比对各组菌在芪归银含药血清干预培养下外膜蛋白的表达谱，从外膜蛋白的差异表达层面明确了芪归银对PA耐药的影响机制。以OPRD2为代表的系列因子表达改变是影响PA菌外膜相关耐药机制的主要基础，亟待进一步深入研究证实。④在上述研究基础上，延伸开展了部分工作：Ⅰ将肺炎治疗的研究中药延伸至清肺消炎丸；Ⅱ结合课题内容，进一步分析了我院2014年临床耐药菌的感染特性，尤其是革兰阴性的PA，革兰阳性的金葡及ICU的菌系分布特点等。在执行该课题过程中，对相应实验器具进行了调整改造，并申获专利三项。