肠道菌群-DAP/NOD1-RIP2信号通路在急性胰腺炎重症化中作用和清下法干预机制

急性胰腺炎（AP）重症化的机制复杂，中医认为AP重症化过程体现了阳明腑实到热毒瘀阻的病机演变。研究表明，肠道菌群在AP重症化中起重要作用，但具体机制不明。我们前期研究显示，AP患者肠道含二氨基庚二酸（DAP）细菌增加，将该细菌移植于动物肠道可加重AP；清下法中药清胰颗粒对AP阳明腑实证有效，且能修复肠道菌群结构、下调NOD1-RIP2炎症通路，这可能与清胰颗粒抑制细菌DAP对其特异性受体NOD1-RIP2通路激活进而阻断炎症级联反应有关。本课题拟采用高通量测序、菌群移植、基因过表达及RNAi等技术，建立高灵敏度质谱定量分析DAP方法，从临床、动物、细胞水平探讨肠道细菌细胞壁成分DAP与NOD1-RIP2通路在AP重症化中的作用机制；阐明清胰颗粒通过抑制细菌DAP/NOD1-RIP2通路防治AP重症化的机理。丰富“清下法”的科学内涵，为AP重症化早期预警和清胰颗粒用药提供理论依据和参考指标。

The mechanism of severe process of acute pancreatitis (AP) is complex. According to Traditional Chinese Medicine, the progression of AP reflects the pathogenesis evolved from “Yangming fu-viscera excess” to “heat toxin stagnation”. Researches have shown that gut microbiota plays an important role in the severe process of AP, but the specific mechanism remains unknown. Our previous studies revealed a significant increase of bacteria containing DAP, which aggravated the condition of AP after being transplanted into the gut of animals. Based on Qingxia therapy, Qingyi Granule, a traditional Chinese medicinal prescription, has proved effective in the treatment of AP with Yangming fu-viscera excess syndrome through repairing the structure of gut microbiota, reducing the expression of NOD1-RIP2 inflammatory pathway and alleviating the inflammatory response. This may be related to the inhibition of bacterial DAP by Qingyi Granule on the activation of its specific receptor NOD1-RIP2 pathway which further blocks the inflammatory cascade reaction. This project aims at employing high-throughput sequencing, bacterial transplantation, gene overexpression, RNAi and other technologies, and establishing the high sensitivity mass spectrometry for quantitative analysis of DAP, to explore the mechanism of bacterial cell wall component DAP and NOD1-RIP2 pathway in the progression of AP at the clinical, animal and cell levels; and to elucidate the mechanism of Qingyi Granule in the prevention and treatment of AP exacerbation by down-regulating bacterial DAP / NOD1-RIP2 pathway. Meanwhile, this project is expected to enrich the scientific connotation of “Qingxia therapy”, and provide theoretical basis and reference index for early warning of severe AP and the clinical application of Qingyi Granule.