4-N-去甲基钩吻素子调控ERS/ER-phagy通路抑制肠道炎症机制研究

钩吻素子具有良好的抗肠道炎症效果。经CYP酶代谢作用，4-N-去甲基钩吻素子的细胞毒性显著降低，并且依旧具有和钩吻素子相近的抗炎药理活性。目前4-N-去甲基钩吻素子抗炎机制研究尚未有报道。因此，本项目以内质网自噬为切入点，采用细胞生物学和分子生物学的方法，构建LPS诱导肠道上皮细胞系IPEC-J2模型，探究内质网应激-内质网自噬信号通路在4-N-去甲基钩吻素子抑制LPS诱导的IPEC-J2炎症中作用。采用组织病理学和分子生物学的方法进一步从体内探究4-N-去甲基钩吻素子抗炎机制。旨在明确4-N-去甲基钩吻素子调控内质网应激-内质网自噬通路增强其抗炎作用。项目创新性地将炎症-内质网应激-内质网自噬三者相互作用的关系等最新研究进展引入4-N-去甲基钩吻素子抗炎作用的机制研究，为钩吻素子和4-N-去甲基钩吻素子的药理作用研究和临床合理应用提供科学依据。

It has confirmed that koumine has a perfect anti-inflammatory effect. With the action of CYP enzyme, the cytotoxicity of N-demethylkoumine, one major metabolite of koumine, is significantly reduced. N-demethylkoumine and koumine have similar anti-inflammatory pharmacological activities. However, the anti-inflammatory mechanism of N-demethylkoumine was still unclear. Thus, endoplasmic reticulum autophagy (ER-phagy), as one key breakthrough point, was focused on the study the anti-inflammatory effect of N-demethylkoumine. Firstly, using the methods of cell and molecular, the mechanisms of endoplasmic reticulum stress (ERS)-endoplasmic reticulum autophagy signaling pathway associated with the preventative effect of N-demethylkoumine on lipopolysaccharide (LPS)-mediated inflammation in IPEC-J2 were investigated. Furtherly, using the methods of histopathology and molecular, the anti-inflammatory mechanisms of N-demethylkoumine were explored. We aimed to find out the significant changes of representative cytokines, of ERS, ER-phagy and inflammation, respectively, to reveal that N-demethylkoumine decreased the productions of pro-inflammatory mediators. Innovatively, the “ERS-ERphagy” signaling pathway was introduced into the anti-inflammatory mechanism of N-demethylkoumine, which may provide the scientific and experimental data reference for pharmacological researches and clinical application of koumine and N-demethylkoumine.