After electroacupuncture treatment at Baihui (DU20) and Qubin (G87), the rat model of middle cerebral artery occlusion (MCAO) was first established. HE, Nissl, and TUNEL staining were used to detect pathological alterations in the rat brain. The relative levels of IL-4, IL-6, TNF-α, and TMAO were determined by ELISA. qRT-PCR and Western blot were used to evaluate the mRNA and protein levels of NF-κB p65, NLRP3, caspase3 and caspase9. Gas chromatography-mass spectrometry (GC-MS) was used to determine the levels of short-chain fatty acids (SCFAs) in rat gut.
Hippocampal cells from rats with spasticity following stroke in the MCAO group were chaotic and loosely distributed with an unclear border, a blurred nucleolus, and vanished cytoplasm when compared to those from the sham operation group. Furthermore, the number of surviving neurons decreased while the number of apoptotic cells increased. In the I/R group, relative levels of IL-6, TNF-α, and TMAO increased considerably, while NF-κB p65, NLRP3, caspase3, and caspase9 were dramatically downregulated. The intestinal contents of n-propyl acetate and propyl butyrate were lowered in rats with spasticity following stroke. Electroacupuncture treatments miraculously remedied all of the foregoing pathogenic alterations.
Pretreatment with electroacupuncture relieves spasticity after stroke by decreasing the inflammatory response, suppressing the NF-κB/NLRP3 signaling pathway, and modulating the gut-brain axis by increasing n-propyl acetate and propyl butyrate levels in the bowel. Our findings establish a new molecular mechanism and theoretical foundation for electroacupuncture therapy of ischemic stroke.
方法:电针治疗百会(DU20)和曲宾(G87)后,首先建立大鼠大脑中动脉闭塞模型(MCAO)。他,Nissl,和TUNEL染色用于检测大鼠大脑的病理改变。IL-4、IL-6、TNF-α的相对水平,通过ELISA测定TMAO。qRT-PCR和Westernblot检测NF-κBp65、NLRP3、caspase3和caspase9的mRNA和蛋白水平。采用气相色谱-质谱法(GC-MS)测定大鼠肠道中短链脂肪酸(SCFA)的含量。
结果:MCAO组脑卒中后痉挛大鼠海马细胞呈混沌、松散分布,边界不清,一个模糊的核仁,与假手术组相比,细胞质消失了。此外,存活的神经元数量减少,而凋亡细胞数量增加。在I/R组中,IL-6、TNF-α、TMAO大幅增加,而NF-κBp65、NLRP3、caspase3和caspase9显著下调。中风后痉挛大鼠的肠道乙酸正丙酯和丁酸丙酯含量降低。电针治疗奇迹般地补救了所有上述致病改变。
结论:电针预处理通过减少炎症反应来缓解中风后的痉挛,抑制NF-κB/NLRP3信号通路,并通过增加肠中乙酸正丙酯和丁酸丙酯水平来调节肠脑轴。本研究为电针治疗缺血性中风建立了新的分子机制和理论基础。