基于LncRNA-miRNA-mRNA调控网络探讨“痰瘀伏邪”病机在糖尿病周围神经病变代谢记忆中的作用及中药干预机制

Long term hyperglycemia could lead to metabolic memory，the intervention which is focus on the controlling of blood glucose is difficult to prevent the progression of diabetic peripheral neuropathy (DPN).How to correctly understand and prevent metabolic memory is important for the prevention and treatment of DPN.However,chronic inflammation is an important factor in the composition of metabolic memory and the microenviromental disorder of peripheral nerve regional immunization inflammation could aggravate the injury of peripheral nerve.Based on our previous study,we put forward that the incubative pathogen prostrate in vivo is similar to the metabolic memory．The pacing factor is viscera disbalance，the principal intergrant are phlegm and blood stasis．In our study,we used animal model of DPN and nervous cells cultured in high glucose to minimic a diabetic condition. The inflammatory immune response as the starting point, many methods were used to investigate the inhibitory effect of Tangzhoutong on inflammatory immune response and immunoinflammation microenviromental disorder of peripheral nerve regional immunization inflammation to relieve the injury of peripheral nerve via metabolic memory. To further investigate the inhibitory effect of Tangzhoutong, the project intends to explore the upstream regulation mechanisms from LncRNA-miRNA-mRNA network,to the downstream cellular immune factor change.More importantly,the protective effect of decoction Tangzhoutong and its molecular mechanisms are also investigated.Our results are both provide a new window and method,but also provide a remarkable evidence in molecular connotation for the therapy of DPN with Tarditional Chinese medicine.

高糖导致的代谢记忆影响下，以控制血糖为重点的干预措施难以阻止糖尿病周围神经病变(DPN)的进展，如何正确认识并阻断代谢记忆是防治DPN的关键。慢性炎症免疫反应是构成代谢记忆的重要因素,局域性炎症微环境紊乱则加重了周围神经损伤。课题组在既往研究基础上从“伏邪理论”着手，认为代谢记忆类似于伏邪在体内伏匿的过程，脏腑功能失调是产生伏邪的基本条件，痰浊、瘀血是伏邪的主要构成部分。研究通过分子生物学和细胞生物学方法，建立高糖代谢记忆动物和细胞模型，以炎症免疫反应为切入点，从整体、细胞、分子水平研究基于痰瘀伏邪论治中药抑制周围神经免疫炎症反应及恢复局域性炎症微环境稳态，从而减轻代谢记忆对周围神经损伤的作用。研究从上游LncRNA-miRNA-mRNA调控网络，到下游相关细胞因子及关键蛋白功能改变，进行微观机制探讨。课题的实施可为相关中药治疗DPN提供理论及实验依据，并为DPN防治提供新的思路和手段。

高糖导致的代谢记忆影响下，以控制血糖为重点的干预措施难以阻止糖尿病周围神经病变的进展，阻断代谢记忆效应是防治糖尿病周围神经病变的关键。慢性炎症免疫反应是构成代谢记忆的重要因素，而局域性炎症微环境紊乱则会明显加重周围神经损伤。课题组在既往研究基础上从“伏邪理论”着手，认为代谢记忆类似于伏邪在体内伏匿的过程，脏腑功能失调是产生伏邪的基本条件，痰浊、瘀血是伏邪的主要构成部分。课题以炎症免疫反应及LncRNA-miRNA-mRNA调控网络为切入点，体内实验以自发性糖尿病GK大鼠作为研究对象，体外实验以原代培养大鼠雪旺细胞为研究对象，建立高糖代谢记忆动物和细胞模型，采用分子生物学、细胞生物学及基因芯片技术，通过LncRNA-miRNA-mRNA的调控作用探讨基于痰瘀伏邪论治中药糖周通抑制周围神经免疫炎症反应及恢复局域性炎症微环境稳态，从而减轻代谢记忆对周围神经损伤的作用机制。研究结果显示，糖周通可以减轻高糖代谢记忆效应所致的周围神经组织及细胞免疫炎症因子表达，改善周围神经组织及细胞病理形态学和超微结构变化。课题组采用基因芯片技术筛选出差异表达基因，根据基因间调控关系，初步构建了LncRNA-miRNA-mRNA基因调控网络。在此基础上采用网络药理学分析了糖周通治疗糖尿病周围神经病变的相关作用靶点，显示糖周通治疗糖尿病周围神经病变的作用涉及多靶点、多条信号通路。本课题的实施为糖周通治疗糖尿病周围神经病变提供了理论及实验依据，并为糖尿病周围神经病变防治提供了新的思路和手段，为研发治疗糖尿病周围神经病变的创新中药奠定了基础。