清热解毒化痰方调控巨噬细胞线粒体自噬/NLRP3干预肠-肺细菌移位治疗重症肺炎的机制研究

重症肺炎死亡率高、严重危害公众健康。肠道屏障、自噬/NLRP3炎症小体活化信号通路在其发病中起着重要作用。基于以往研究，进一步探讨清热解毒化痰方治疗本病的机制，拟开展研究：①制备重症肺炎大鼠模型，采用病理学、ELISA、免疫组化等技术观察肺泡灌洗液和肺组织中的细胞分类、炎症因子及组织病理形态学；采用免疫组化、ELISA、qPCR、WB、荧光标记等技术，从线粒体自噬、NLRP3基因与蛋白表达、炎症因子、肠道通透性、细菌移位等方面观察中药治疗重症肺炎大鼠的作用机制。②以巨噬细胞/肠道上皮细胞为研究对象，采用细胞共培养、血清药理学、Seahorse、qPCR和WB等技术，从细胞吞噬能力、线粒体功能、线粒体自噬、细胞增殖与凋亡、NLRP3活化、肠上皮细胞屏障功能等方面探讨清热解毒化痰方调控自噬/NLRP3活化治疗重症肺炎的机制。以揭示中医药治疗重症肺炎机制，为中医临床应用和新药研发提供依据。

Severe Community acquired pneumonia has a high mortality rate and seriously endangers public health. Intestinal barrier and autophagy / NLRP3 inflammasome activation play an important role in its pathogenesis. On the basis of previous studies, to explore the mechanism of Qingre jiedu Huatan recipe in the treatment of this disease, to carry out: (1) to establish a rat model of severe Community acquired pneumonia, observe the cell classification, inflammatory factors and histopathology in bronchoalveolar lavage fluid and lung tissue by pathology, ELISA and immunohistochemistry; Immunohistochemistry, ELISA, qPCR, WB and fluorescence labeling techniques were used to observe the mechanism of traditional Chinese medicine in treating rats with severe Community acquired pneumonia from the aspects of mitochondrial autophagy, expression of NLRP3 gene and protein, inflammatory factors, intestinal permeability and bacterial translocation. (2) To explore the mechanism of Qingre jiedu Huatan recipe in regulating autophagy / NLRP3 activation in the treatment of severe pneumonia from the aspects of cell phagocytosis, mitochondrial function, mitochondrial autophagy, cell proliferation and apoptosis, NLRP3 inflammasome activation, intestinal epithelial cell barrier function, etc., using the techniques of cell co-culture, serum pharmacology, Seahorse, qPCR and WB, etc. to explore the mechanism of Qingre jiedu Huatan recipe in regulating autophagy / NLRP3 inflammasome activation in the treatment of severe pneumonia. In order to reveal the mechanism of traditional Chinese medicine in the treatment of severe pneumonia and provide a reference for clinical application and new drug research and development of traditional Chinese medicine.