LITA-VNS调控神经免疫治疗房颤机制及与心肌外泌体关联性研究

Atrial fibrillation (AF) is one of the major cardiovascular disorders with higher morbidity and disability as well poor treatment.effect.Inflammation and cardiac autonomic nerve dysfunction are the important pathogenesis of AF.The treatment effectiveness of AF has been improved significantly by β-blockers,cervical vagus nerve stimulation(VNS),cardiac plexus(GPs)ablation as well renal denervation(RD).A recent study from basic research shows that Low-Level transcutaneous auricular vagus nerve stimulation（LITA-VNS）is effective in treatment of AF induced by atrial pacing .Safety,effective as well side effects of the new method and the underlying mechanisms are needed to be clarify.The molecular mechanism of exosomes in the transmission of information between nerve-immune cells are leading subject。Based on our previous studies，we proposed a hypothesis: Myocardial exosomes transmit information between CANS and "cholinergic anti-inflammatory pathway" in LITA-VNS treatment of AF。In this project，we will produce a canine AF model by 4 weeks atrial pacing.With implementation of LITA-VNS, we will monitor AF inducibility,nerve activity in VNT、SG and GPs, atrial electrol、structure and nerve remodeling, cholinergic nerve anti-inflammatory pathway as well as cardiac exosomes before and after cervical vagus nerve stimulation or ablation by cross tracing neurons with a virus, neural signal recording and molecular biology technology .We will investigate the optimal stimulation intensity、frequency and location of LITA-VNS inhibition in AF. The influence of LITA-VNS on AF inducibility 、atrial electrical 、structure remodeling and neuroplasticity、inflammatory factors as well myocardial exosomes level will be evaluated. The influence of LITA-VNS on regulation of autonomic nerve system and cholinergic anti-inflammatory pathway as well relationship with cardiac exsomes will be analyzed. Finally,we will clarify the neroinflammation mechanism and the function of exosome in transmission of information between nerve-immune cells in order to provide better theoretical basis for transforming LITA-VNS to clinic application for treatment of AF.

心脏自主神经功能（CANS）紊乱和炎症是房颤（AF）重要的致病机制。新近研究发现：低强度电压刺激耳屏迷走神经支（LITA-VNS）是治疗AF的新途径,但方法学未建立且机制不明。外泌体在神经免疫细胞间传递信息分子机制的研究是前沿课题。本项目依据前期研究基础并查阅文献提出假设：LITA-VNS通过调控CNAS和抗炎机制治疗AF，心肌源外泌体参与了调控的信息传递。本课题通过制作犬AF模型，用病毒跨神经元示踪、神经信号记录、MEA和分子生物学等技术，研究LITA-VNS刺激的强度、频度和部位变化对AF的影响差异，分析外泌体miRNA-133a和miRNA-1变化与LITA-VNS调控CNAS影响AF诱发率、心房肌电结构和神经重构、炎性因子等变化的关系。建立LITA-VNS治疗AF的有效方法学，并阐明其调控神经免疫的机制及与外泌体的关联性，为LITA-VNS临床转化治疗AF提供理论依据。

心脏自主神经功能（CANS）紊乱和炎症是房颤（AF）重要的致病机制。新近研究发现：低强度电压刺激耳屏迷走神经支（LITA-VNS）是治疗AF的新途径,但方法学未建立且机制不明。外泌体在神经免疫细胞间传递信息分子机制的研究是前沿课题。本项目依据前期研究基础并查阅文献提出假设：LITA-VNS通过调控CNAS和抗炎机制治疗AF，心肌源外泌体参与了调控的信息传递。本课题通过制作犬AF模型，用病毒跨神经元示踪、神经信号记录、MEA和分子生物学等技术，研究LITA-VNS刺激的强度、频度和部位变化对AF的影响差异，分析LITA-VNS调控CNAS影响AF诱发率、心房肌电结构和神经重构、炎性因子等变化的关系。建立LITA-VNS治疗AF的有效方法学，并阐明其调控神经免疫的机制及与外泌体的关联性，为LITA-VNS临床转化治疗AF提供理论依据。本研究通过制作单纯颈部迷走神经刺激介导的犬房颤模型，研究迷走神经刺激时间和心房起搏频率依赖对房颤诱发率的影响。主要研究结果：（1）完成犬心房起搏 4 周 AF 模型的制作，确定 LIT-VNS 治疗 AF 的最佳刺激参数。（2）探讨神经刺激仪对每个麻醉后大鼠的耳屏、耳甲、耳轮分别进行电刺激LL-VNS治疗房颤的最佳刺激部位、最佳刺激强度以及神经机制。（3）探讨经皮迷走神经刺激耳甲和耳轮对迷走神经放电的影响。（4）HL-1小鼠心肌细胞信号检测及房颤模型建立，以及外泌体的分离表征鉴定。等等。