基于ceRNA网络探讨电针干预TLR4/Sphk1/S1P信号通路调控CIS后血管新生的机制研究

针对缺血性脑卒中高发病率、高致残率、高复发率和高死亡率特点，临床上电针头穴治疗的有效性已被认可，但电针调控CIS后血管新生以减轻神经损伤的作用机制仍未明了。前期研究发现脑缺血AG启动与TLR4基因介导炎症级联反应调控Sphk1-S1P/S1PR1途径以作用血管内皮功能密切相关。因此我们提出“电针头穴启动了ceRNA网络效应，刺激lncRNA表达，同时内源性竞争负调控靶miRNA，促进TLR4mRNA表达，激活Sphk1-S1P/S1PR1信号通路，纠正血管内皮功能紊乱以促CIS后血管新生以发挥脑保护作用”的假说，构建MCAO模型，利用基因测序、慢病毒转染、real-time PCR、Western blot等技术，研究TLR4基因在电针治疗CIS过程中影响血管内皮细胞增殖及凋亡的分子机制，为明确电针对CIS的治疗效应提供可靠的依据。

The effectiveness of electro-acupuncture treatment for cerebral ischemic stroke (CIS), which is characterized by its high incidence, high disability rate, high recurrence and high mortality rate, has been approved in clinical practices, while the mechanism of electro-acupuncture in regulating angiogenesis by alleviating nerve injury remains unclear.Previous studies showed that AG activation in cerebral ischemia is closely linked to the regulation of Sphk1-S1P/S1PR1 pathway by the inflammatory immune cascade mediated by TLR4 gene to play a role in vascular endothelial function. Therefore, we propose the hypothesis that electro-acupuncture may activate ceRNA network, stimulate lncRNA expression, and interfere negative regulation target miRNA, promote TLR4 mRNA expression, activate Sphk1-S1P/S1PR1 signaling pathway, and correct endothelial dysfunction by angiogenesis to exert a protective effect. Thus, MCAO model will be constructed, high-throughput sequencing, Lentivirus transfection, real-time PCR, western blot and other techniques will be used to study the molecular mechanism of vascular endothelial cell proliferation and apoptosis affected by TLR4 gene during the electro-acupuncture treatment of CIS, and to enrich the effect of electro-acupuncture treatment of CIS angiogenesis.