痰瘀同治法对心肌缺血再灌注损伤中核转录因子-κB与自噬交互作用的调控机制研究

NF-κB and autophagy are activated and involved the process of regulation of immunity, inflammation, growth control and cell apoptosis during reperfusion injury. In addition, NF-κB and autophagy have a common upstream signal and regulator, mutual control should be exerted through the positive or negative feedback effect, thus ensuring the steady-state response at the molecular level.Therefore，clarify the relationship between NF-κB and autophagy will creates a new way for decode the complex mechanism of MI/RI..With the helping of the early Nation Nature Science Fund，the experiment confirmed that NF-κB signal transduction pathway could be activated in the process of MI/RI. The treatment of phlegm and bloood stasis resolved simultaneously could protect myocardium from inflammatory damage, its mechanism might restrain the activation of upstream kinase of NF-κB signal transduction pathway. And based on this conclusion, this research focus on the interaction between NF-κB and autophagy and analysis the adjustment mechanism, study the effect of this mechanism on myocyte apoptosis and inflammatory response in MI/RI. Moreover,the therapy with phlegm and bloood stasis resolved simultaneously will be the means of intervention.The accommodation mechanism will be revealed partly through observing the effect of the treatment on the specific sites of NF-κB signal transducer pathway and autophagy in the process of MI/RI.The study will provide Chinese treatment of MI/RI with innovative ideas and experimental evidence.

自噬与NF-κB的激活均可参与再灌注后的免疫调节、炎症反应、生长控制及细胞凋亡的过程，而且，在分子水平上，自噬与NF-κB有着共同上游信号和调节器，两者通过积极的或负反馈回路而相互控制，从而保证稳态反应。因此，阐明自噬和NF-κB之间的相互关系将为译解复杂的缺血再灌注损伤发生机制开阔新思路。.在前期国家自然科学基金资助下，已证实MI/RI可激活NF-κB信号转导通路且痰瘀同治法可通过抑制该通路达到抗炎、减轻心肌损伤的作用。因此，本研究将在此基础上，以NF-κB与自噬反应交互作用为切入点，深入探讨两者在MI/RI过程中的相互调节机制，及其对再灌注后心肌炎症、凋亡程度的影响；同时以痰瘀同治方为干预手段，重点观察该方对自噬、NF-κB通路的具体调节位点，并试图揭示痰瘀同治方在两者交互关系中的作用，力求从一崭新的角度阐释痰瘀同治方对MI/RI的作用机理；同时也为优化治疗策略提供创新策略和依据。

本研究在前期工作基础上，以NF-κB与自噬反应交互作用为切入点，深入探讨两者在心肌缺血再灌注损伤过程中的相互调节机制，及其对再灌注后心肌炎症、凋亡程度的影响；同时以痰瘀同治方为干预手段，重点观察该方对自噬、NF-κB通路的具体调节位点，揭示了痰瘀同治法在两者交互关系中的作用及其对心肌缺血再灌注损伤的作用机理。. 实验显示，心肌缺血再灌注损伤可同时激活NF-κB转录和自噬过表达， NF-κB的活化可以促使自噬反应加强，而再灌注后自噬的强烈反应也可刺激NF-κB活性的进一步升高。而且，再灌注后心肌细胞凋亡及炎症反应的发生、发展也与自噬及NF-κB的活化密切相关，凋亡及炎症损伤的程度可随着自噬及NF-κB活性的加强而愈加严重。由此可见，自噬与NF-κB通路两者间可以通过关键元件如NF-κB、Beclin-1等相互促进，共同协作，引发凋亡、炎症等一系列损伤反应而导致再灌注损伤的进一步恶化。结果证实，痰瘀同治法可以通过抑制NF-κB活化、调控自噬基因Beclin-1、LC3的过表达，阻断两者的交互协同影响，通过多种途径、多靶点起到改善心肌细胞凋亡，减轻炎症反应的抗再灌注损伤心肌保护作用。. 研究结果从NF-κB与自噬关系的角度阐释了痰瘀同治法抗心肌缺血再灌注损伤的作用机理，为中医药治疗缺血性心脏病提供了创新性思路和实验依据。