基于糖酵解途径探讨清宣通络方改善肺炎支原体对大环内酯类抗生素耐药性的机制研究

随着大环内酯类耐药肺炎支原体（MRMP）感染发病率迅速增加，如何应对MRMP感染成为临床研究的一大挑战。近年来发现病原菌可以通过提高自身能量代谢水平来提高对抗生素的抗性，糖酵解途径在其中起重要作用。课题组前期研究已证实，清宣通络方在治疗MPP时能增强抗生素敏感性，提高退热疗效，明显缩短病程，在调节免疫炎症损伤中与阿奇霉素具有同等效力，且可通过提高宿主的糖酵解代谢产能，多靶点，全方位调节人体内环境。预实验结果显示清宣通络方含药血清能改善阿奇霉素诱导的MP耐药。本研究拟以体外诱导MP耐药菌株为切入点，以“能量代谢中糖酵解水平变化能够影响MP对大环内酯类抗生素耐药性”为主线，采用质粒转染、阿奇霉素蓄积实验、HE染色、流式细胞术等实验技术，从组织形态、细胞、分子生物学等不同层面探讨清宣通络方改善肺炎支原体对大环内酯类抗生素耐药性的作用机制，为中医药改善抗生素耐药提供可靠的实验室依据。

With the rapid increase in the incidence of macrolide resistant mycoplasma pneumoniae (MRMP) infection, how to deal with MRMP infection has become a major challenge in clinical research. In recent years, it has been found that pathogenic bacteria can improve their resistance to antibiotics by improving their own energy metabolism, in which glycolysis pathway plays an important role. The previous study of the research group has confirmed that Qingxuan Tongluo formula can enhance the sensitivity of antibiotics, improve the antipyretic effect, shorten the course of disease obviously, have the same effect as azithromycin in regulating immune inflammatory injury, and can regulate the internal environment of human body in an all-round way by improving the capacity of glycolysis and metabolism of the host and multi-targets. The pre-experimental results showed that the drug-containing serum of Qingxuan Tongluo formula could improve the MP resistance induced by azithromycin. The purpose of this study is to induce MP-resistant strains in vitro as the starting point, with "the change of glycolysis level in energy metabolism can affect the resistance of MP to macrolides" as the main line, using plasmid transfection, azithromycin accumulation test, HE staining, flow cytometry and other experimental techniques. To explore the mechanism of Qingxuan Tongluo formula in improving the resistance of mycoplasma pneumoniae to macrolide antibiotics from different aspects of histomorphology, cell and molecular biology, so as to provide reliable laboratory basis for traditional Chinese medicine to improve antibiotic resistance.