基于NLRP3炎症小体信号通路的桦褐孔菌多糖的抗炎机制及对结肠炎相关结直肠癌的影响

The polysaccharide from inonotus oblique has anti-inflammatory and anti-tumor effects. However, the mechanism of anti inflammation is related to the NLRP3 inflammatory pathway, and the effect of colorectal cancer (CRC) is rarely studied. In particular, have not been seen in polysaccharides of inonotus oblique on colitis associated colorectal cancer (CAC) research reports. Therefore, the purpose of this study is to clarify the inonotus oblique polysaccharide on anti inflammatory mechanism, LPS pretreated mice macrophages and mouse bone marrow derived dendritic cells/macrophages, were treated with polysaccharide of inonotus oblique, the proinflammatory cytokines were checked by ELISA in the detection of cells, such as TNF-alpha, IL-1 beta, IL-6, IL-18, pro- IL-1 beta secretion; The activation of NF - kapa B and the expression of COX-2, iNOS, NLRP3, ASC, IL-18, caspase-1 were detected by western blot assay. In order to elucidate effects of the polysaccharides from inonotus oblique in CAC and its mechanisms, polysaccharide from inonotus oblique AOM/DSS -induced mice models of CAC were observed in mice bowel inflammation and cancer and immunological changes. Primary colorectal cancer cells were isolated and cultured from the tissue samples of patients undergoing CRC operation. To observe the anti-tumor effect of nonotus oblique polysaccharide on this cell, and to detect the expression of NLRP3 inflammatory body protein in serum and tissue of patients (IBD and CRC ). For from molecular and cell level discussion its role mechanism, with birch brown hole bacteria polysaccharide stimulus colon cancer strains, and small rat giant bite cell strains and the original generation giant bite cell and intestinal lamina propria mononuclaer cell (LPMC), and intestinal epithelial cell, using molecular biology, and protein chip technology, and immune imprinted, immunochemical technology, observe pro-inflammatory cytokines and inflammatory factors (COX-2) and its signal pathway, while observe NLRP3 inflammasome pathway. This study will provide polysaccharide from inonotus obliquus or other polysaccharides of traditional Chinese medicine prevention and cure other inflammation-associated cancer research. It also provides new targets for the diagnosis and treatment of IBD and CRC patients.

桦褐孔菌多糖具有抗炎、抗肿瘤等作用，但其抗炎作用机制是否与NLRP3炎症小体通路有关，并对结直肠癌效应的研究很少，尤其尚未见到该多糖对结肠炎相关结直肠癌（CAC）的效应及其机制方面的研究报道。因此，本研究为了阐明该多糖抗炎作用机制，用该多糖处理炎症条件下的小鼠巨噬细胞等后，利用ELISA法检测促炎细胞因子的分泌，利用免疫印迹等技术检测NLRP3-ASC-caspase-1炎症小体，沉默炎症小体蛋白和NF-kB后，检测IL-18等及iNOS和COX-2的表达；为探讨对CAC的影响，将该多糖供给AOM/DSS诱导的CAC小鼠模型，观察不同时期小鼠肠道炎症程度及癌变程度，用结肠癌细胞株探讨其机制，从临床组织标本中分离培养原代结直肠癌细胞后，观察该多糖的抗肿瘤效应；在临床IBD和结直肠癌患者的血清和组织标本中检测NLRP3炎症小体等。本研究将为用该多糖防治炎症相关肿瘤的研究方面提供理论依据。

桦褐孔菌多糖具有抗炎、抗肿瘤等作用，但其抗炎作用机制是否与NLRP3炎症小体通路有关，并对结直肠癌效应的研究很少，尤其尚未见到该多糖对结肠炎相关结直肠癌（CAC）的效应及其机制方面的研究报道。因此，本研究首先阐明了桦褐孔菌多糖可通过调控NLRP3炎症小体的激活，控制炎症反应，并阐明了NLPR3炎症小体与IBD及CRC疾病之间的相关性。其次探讨了桦褐孔菌多糖对CAC具有抑制癌变作用，尤其是NLRP3炎症小体的具体作用机制，为利用桦褐孔菌多糖预防和治疗其他炎症性疾病提供依据，为长白山地区桦褐孔菌的开发提供新的理论依据，也为研究其他中药材多糖物质的抗炎、抗肿瘤作用机制方面提供方法和技术思路。