扶正透邪、解毒化瘀法对多重耐药铜绿假单胞菌的抑菌作用及免疫损伤的干预机制研究

Multi-drug resistant pseudomonas aeruginosa, the most common conditional pathogen in clinic, will be used to make a senile immune injury mice model through abdominal innoculation in this study. Right-reinforcing pathogen-dispersing toxin-stasis removing formula that have been proven effective in previous studies plus qi-tonifying pathogen-dispersing herbs based on clinical experiences and other experimental studies are given to the treatment group. We aim to observe the in vitro antibacterial efffects of this formula in decoction preparationa by using disk diffusion and tube dilution treatment methods and by observing the ultrastructure changes through electromicroscope compared with those of medicated mice serum. We also will evaluate the dynamic levels of cytokines at different phases and measure the pathological changes of greater omentum, lung, thymus, and spleen tissue so as to explore the antibacterial and immune injury intervention mechanism of right- reinforcing pathogen-dispersing toxin-stasis removing method for multi-drug resistance pseudomonas aeruginosa. This is a pragmatic study of high value for the screening of effective antibacterial herbal therapy or even antagonists of drug-resistant bacteria to enhance the effectiveness of antibotics and reduce its dosage. This study will provide further evidence for the efficacy of this formula in the treatment of drug-resistant senile pneumonia caused by the most common conditional pathogen of multi-drug resistant pseudomonas aeruginosa. It will also act as references of methodological approach to the development of TCM febrile theories.

以目前临床上最常见的条件致病菌之一的多重耐药铜绿假单胞菌为研究对象，并以该耐药菌腹腔感染所致老年大鼠免疫炎性损伤为试验模型，选择在前期研究中发现治疗老年人耐药菌肺炎疗效显著的扶正解毒化瘀治法方药，并根据临床经验及前期的实验研究结果加用益气透邪类中药，通过采用纸片扩散法、试管稀释法，并运用电子显微镜观察细菌的超微结构，探讨扶正透邪、解毒化瘀治法方药的水煎剂、含药血清对多重耐药铜绿假单胞菌的体外抑制作用，同时评价该治法方药对试验动物模型不同时相炎性细胞因子的水平及大网膜、肺、胸腺、脾脏等器官组织病理改变的影响，阐明该治法方药对多重耐药铜绿假单胞菌的抑菌作用及免疫损伤的干预机制。本课题针对日益严重的细菌耐药这一难题开展上述研究，为临床指导拓宽使用有效的中药抑菌剂或耐药菌的逆转剂，并增强抗菌药物的疗效，进而减轻多重耐药菌的用药压力，具有重要的现实意义，也为中医热病学术的发展提供可借鉴的方法学内容。

铜绿假单胞菌是院内感染的重要病原菌，其感染率高、耐药性强、致死率高，严重威胁患者的健康。抗生素的大量广泛应用，使细菌耐药问题日趋严峻。老年人免疫功能低下，基础疾病较多，往往发生耐药菌感染，这是临床中非常棘手的问题。本课题以前期工作中发现治疗老年人耐药菌肺炎疗效显著的扶正解毒化瘀方为基础，加用实验研究中发现具有体外抑菌作用及调节机体免疫功能的扶正透邪中药，组成扶正透邪解毒化瘀治法方药，以多重耐药铜绿假单胞菌为研究对象，通过体外抑菌实验，发现扶正透邪解毒化瘀治法方药联合亚胺培南、头孢他啶均具有协同抑菌作用；通过动物实验，发现中西医结合治疗组可明显降低大鼠肺组织内促炎因子TNF-α、IL-1β、IL-6、IL-2、IFN-γ的释放，其疗效优于单纯西药组，中药提前给药组在炎症反应早期可以升高抗炎因子IL-4、IL-10的含量，在多重耐药铜绿假单胞菌感染后最严重的3-5天能明显降低促炎细胞因子TNF-α、IL-1β、IL-6、IL-2、IFN-γ的表达，基于此结果，我们考虑扶正透邪解毒化瘀治法方药可能通过抑制促炎因子的过度释放、提高抗炎因子的表达，调节机体在多重耐药菌感染状态下失衡的炎症因子趋向平衡状态,从而减轻机体过度的炎症反应，促进组织器官的修复，最终降低了多重耐药铜绿假单胞菌肺炎大鼠的死亡率。本课题在为临床拓宽使用有效的中药抑菌剂或耐药菌的逆转剂、增强抗菌药物的疗效、减轻多重耐药菌的用药压力方面，具有重要的现实意义，也为中医热病学术的发展提供实践方法学内容。