基于MicroRNA-内质网应激-自噬轴探讨益气温阳活血利水法治疗心衰的机制

The incidence of heart failure is steadily increasing. The important role of autophagy in heart failure has been paid much attention. To explore the mechanism of autophagy in heart failure will contribute to find the upstream targets to treatment heart failure. Recent studies demonstrated that both miRNA and endoplasmic reticulum stress (ER stress) had played an importannt role in HF. The miRNA -ERS axis mediated cardiomyocyte autophagy may play a significant role in HF. Therapies targeted against miRNA-ER stress-autophagy axis may become an imporant strategy in patients with HF. Preliminary study showed that the thearpy of benefiting qi and warming yang and activating blood can regulate cardiomyocyte autophagy and improve cardiac function. This research will explore the mechanism of miRNA-ER stress-autophagy axis in vivo and in vitro by using the different methods , such as Western Blot, qPCR, adenovirus vector transfection and transmission electron microscope(TEM) on the overall animal models and primary cells. The results of the research will provide new theoretical basis and experimental evidence for treating heart failure with benefiting qi and warming yang, activating blood and promoting diuresis. It can provide evidence on screening new traditional Chinese medicine compounds that can regulate the miRNA-ER stress-autophagy axis in heart failure.

心衰的发病率稳步增加，自噬在心衰发病中的重要作用已开始受到广泛重视，深入探索自噬在心衰中作用的上游机制将有助于寻找到从自噬途径治疗心衰的上游靶点。最新研究证明miRNA和内质网应激在心衰的发病中均起着重要作用，miRNA-内质网应激-自噬轴在心衰的发病中发挥着重要作用，调节miRNA-内质网应激-自噬轴成为心衰防治的重要策略。益气温阳活血利水法是心衰的主要治法，研究证明其具有调节心肌细胞自噬，改善心功能等作用。本研究拟通过制备整体动物模型和培养原代细胞，利用免疫印迹、qPCR、腺病毒载体转染、透射电镜等方法，在细胞和整体水平研究心肌细胞miRNA-内质网应激-自噬轴在心衰的作用及机制，以及益气温阳活血利水法调节miRNA-内质网应激-自噬轴治疗心衰的作用。为益气温阳活血利水法治疗心衰提供新的理论依据和实验基础，为筛选调节miRNA-内质网应激-自噬轴治疗心衰的中药新药提供依据。

心衰的发病率稳步增加，自噬在心衰发病中的重要作用已经受到广泛重视，研究证明miRNA和内质网应激在心衰的发病中起着重要作用，miRNA-内质网应激-自噬可能在心衰的发病中发挥着重要作用，调节miRNA-内质网应激-自噬轴可能成为心衰防治的重要策略。益气温阳活血利水法作为心衰防治的主要治法，可能通过调节miRNA-内质网应激-自噬轴防治心衰。本研究在动物和细胞水平探讨内质网应激-自噬轴在心衰中的作用，以及miRNA在心衰时对内质网应激-自噬轴的调节作用。并深入探讨益气温阳活血利水法对心衰心肌细胞miRNA-内质网应激-自噬轴的调节作用。本项目按照计划开展研究，研究目标完成良好。在动物整体水平发现了miRNA221、miRNA214内质网应激-自噬轴介导了心衰时心肌细胞自噬，抑制miRNA221或miRNA214均能通过抑制内质网应激介导的自噬，改善慢性心衰心慢性心衰心功能和心肌肥大。益气温阳活血代表方心阳片低中高剂量能有效改善慢性心力衰竭心功能。结果提示miRNA221、miRNA214-内质网应激-自噬轴介导了心肌细胞自噬在慢性心衰发病中发挥着重要作用，益气温阳活血代表方心阳片中高剂量具有调节miRNA221、miRNA214-内质网应激-自噬轴防治慢性心力衰竭的作用。在细胞水平，证明了血管紧张素II刺激可导致大鼠乳鼠心肌细胞内质网应激和自噬增加强，并导致心肌细胞肥大。抑制miRNA221、miRNA214可减轻AngII刺激导致的心肌细胞内质网应激和自噬增强，并抑制心肌细胞肥大。益气温阳活血代表方心阳片含药血清能有效抑制AngII刺激导致的心肌细胞内质网应激和自噬的增强，并改善AngII刺激导致的心肌细胞肥大。结果提示AngII刺激下，miRNA221、miRNA214-内质网应激-自噬轴导致了心肌细胞肥大，益气温阳活血代表方心阳片含药血清具有调节miRNA221、miRNA214-内质网应激-自噬轴减少AngII刺激下心肌细胞肥大的作用。该研究结果为未来筛选治疗慢性心力衰竭药物提供了实验基础。同时为益气温阳活血法及其代表方心阳片未来在临床的推广应用提供了实验依据和理论支持，也将为益气温阳活血法及其代表方心阳片广泛治疗慢性心力衰竭带来较好的社会效益和经济效益。