有氧运动通过NF-κB通路调控自噬改善动脉粥样硬化的作用研究

Atherosclerosis (AS) is pathological basis of coronary heart disease and many cardiovascular diseases. It has been confirmed that Long-term regular exercise training can inhibit AS. Studies have shown that NF-κB and autophagy play an important role in the occurrence and development of AS. And NF-κB pathway regulating autophagy has been studied in other areas, however, whether the NF-κB signaling pathway involved in the regulation of autophagy in AS has not been reported. Based on this, we proposed the following hypothesis: NF-κB signaling pathway mediated autophagy may be an important mechanism of aerobic exercise to prevent or delay the formation of AS. Multidisciplinary techniques and methods were used in present study. Firstly, the rat model of atherosclerosis was established, and the related protein expression of NF-κB and autophagy in different groups were compared, clarifying the the correlation between NF-κB and autophagy in mechanism of aerobic exercise on AS, confirmed mediated effects of NF-κB on autophagy at the animal level. Then the AS rat arterial endothelial cells were cultured by NF-κB specific agonists and antagonists. The expression of NF-κB and autophagy were compared, further exploring the relationship between NF-κB and autophagy at the cell level.

动脉粥样硬化（AS）是冠心病及众多心血管疾病的病理基础。长期有规律的运动训练对AS的预防和延缓作用已经证实，然而其机制尚不完全清楚。研究显示，NF-κB和自噬均在其中发挥重要作用。其它领域研究显示NF-κB通路对细胞自噬具有调控作用。然而，NF-κB信号通路调控细胞自噬是否参与了有氧运动对AS的作用尚未见报道。基于此，提出如下假设：NF-κB信号通路调控自噬可能是运动改善血管内皮功能，延缓AS的重要作用机制之一。本研究拟运用多学科的技术与方法，首先构建大鼠AS模型，通过对比不同干预措施下NF-κB和自噬相关基因的表达情况，阐明NF-κB与自噬之间的相关性，从动物层面证实NF-κB对自噬的调控作用。然后，培养AS鼠动脉内皮细胞，通过对比不同处理措施间NF-κB和自噬的表达，从细胞层面证实NF-κB对自噬的调控作用。预期研究结果对于明确有氧运动在冠心病康复中的作用机制具有重要意义。

动脉粥样硬化（AS）是冠心病及众多心血管疾病的病理基础。长期有规律的运动训练对AS的预防和延缓作用机制尚不完全清楚。课题组在以往研究的基础上，提出了NF-κB调控自噬可能是其重要机制的假设。基于此，本研究从动物和细胞两个层面阐述了NF-κB调控自噬在有氧运动改善AS机制中的作用。动物实验部分，根据干预措施不同分为有氧运动组、模型组和PDTC组，结果表明，有氧运动组的TC、TG和Ox-LDL均显著低于模型组，表明有氧运动可有效降低血脂水平，同时病理组织学观察也发现有氧运动可以有效延缓斑块的形成；有氧运动组和PDTC组胞浆NF-κB p65的表达均显著高于模型组，而细胞核NF-κB p65的表达均低于模型组，表明有氧运动能够下调AS形成过程中的NF-κB水平；有氧训练组和 PDTC 组 Beclin1和 LC3B II/I的表达水平明显高于对照组，IF染色也显示，两组的LC3BⅠ/I阳性表达也高于模型组，提示有氧训练和PDTC 均能激活细胞自噬，同时表明NF-κB下调可能对自噬具有激活作用。在细胞层面，选取AS 鼠HUVEC进行培养，并根据处理方法不同随机分为七组。EMSA检测和Western blot分析均表明，Ox-LDL和 PMA可以激活 NF-κB 的表达，siRNA 可以抑制NF-κB的表达。Beclin1和LC3BII/I的检测结果说明，Ox-LDL和PMA可以抑制自噬，PDTC和p65 siRNA可以激活细胞自噬。MDC结果同样表明Ox-LDL和PMA可以抑制自噬，而PDTC可以激活自噬，即NF-kB下调能够上调HUVEC自噬水平。总之，本研究通过从动物和细胞两个层面，证实了有氧运动下调NF-κB进而激活自噬是有氧运动改善动脉粥样硬化的重要机制之一。研究结果对于有氧运动在心肺康复中的推广开展提供了科学依据，同时也为其它途径调控自噬提供新的思路。