基于NLRP3/STAT3信号通路介导的焦亡-自噬Crosstalk的夹脊电针促进脊髓损伤修复的神经元再生机制

The neuronal regeneration (NR) plays a pivotal role on the recovery after spinal cord injury (SCI). Our previous studies have illustrated that Jiaji electroacupuncture (JJEA) improved the recovery after SCI through regulation of antioxidation induced by Nrf2. Additionally, the recent researches have indicated that nuclear factor erythroid-2 related factor2 (Nrf2) can regulate pyroptosis and autophagy (PA), and both STAT3 signaling pathway induced autophagy and NLRP3 signaling pathway induced pyroptosis are involved in NR. Whereas, the detailed mechanisms of the crosstalk between PA in neuronal regeneration are yet to be elucidated. Thus, Allen’s SCI rat models will be established in this project, and test of BBB, Nissl, WB, PCR, IF etc. will be used to observe the locomotor function, expression of cells, levels of protein and mRNA of factors related of PA and NR. Which will be claimed by this project is that the recovery effect of JJEA treating SCI based on pyroptosis induced by NLRP3 and autophagy induced by STAT3 respectively. Furthermore, it will clearly define about the mechanisms of JJEA promoting NR after SCI based on PA crosstalk induced by the NLRP3/STAT3 signaling pathway. Furthermore, this project will analyze the mechanisms of JJEA promoting restoration after SCI scientifically and provide theoretical evidence for promoting further spread of JJEA treating SCI in clinic.

神经元再生(NR)是脊髓损伤(SCI)修复的关键机制。课题组前期已证实，夹脊电针通过调控Nrf2介导的抗氧化反应可以促进SCI修复。此外，已有研究表明Nrf2可调控焦亡-自噬(PA)，并且STAT3介导的自噬、NLRP3介导的焦亡均参与了NR。但是，PA crosstalk在NR中的详细机制尚不明确。因此，本项目拟应用Allen’s SCI模型，采用BBB、Nissl、WB、PCR、IF等方法，观察夹脊电针对SCI大鼠的运动功能、细胞表达、PA和NR相关因子的蛋白及mRNA水平的影响。从整体、细胞及分子水平明确夹脊电针分别基于NLRP3介导的焦亡和STAT3介导的自噬对SCI的修复作用，阐明夹脊电针基于NLRP3/STAT3信号通路介导的PA crosstalk促进SCI的NR机制，进而科学解析夹脊电针修复SCI的内涵，为进一步临床推广夹脊电针治疗SCI提供理论依据。

前期研究表明Nrf2可调控焦亡-自噬(PA)，并且STAT3介导的自噬、NLRP3介导的焦亡均参与了神经元再生(NR)。PA crosstalk在NR中的详细机制尚不明确。因此，本项目通过对大鼠脊髓损伤（SCI）发生及夹脊电针治疗后不同时间点进行BBB评分，HE染色、Nissl染色、Elisa、Western blot、RT-PCR、IF等实验检测手段，观察夹脊电针对SCI大鼠的运动功能、细胞表达、PA和NR信号通路中标志性信号分子的蛋白及mRNA水平的影响，从功能、结构和细胞分子信号多维度系统剖析。结果提示，夹脊电针可改善SCI大鼠后肢运动功能、组织损伤情况、炎性细胞浸润，可调控PA crosstalk中标志性信号分子蛋白及mRNA的表达。结论认为夹脊电针可基于NLRP3介导的焦亡和STAT3介导的自噬对SCI的修复作用，阐明夹脊电针基于NLRP3/STAT3信号通路介导的PA crosstalk促进SCI的NR机制，为夹脊电针治疗SCI提供理论及实验依据。