Electroacupuncture pretreatment alleviates spasticity after stroke in rats by inducing the NF-κB/NLRP3 signaling pathway and the gut-brain axis.

OBJECTIVE: Spasticity is one of the most prevalent ischemic stroke sequelae and the leading cause of disability after stroke. Although electroacupuncture pretreatment has been shown to be effective in the treatment of ischemic stroke, its therapeutic effect and mechanism on post-stroke spasm remain unknown. The purpose of this study was to look into the potential mechanism of electroacupuncture pretreatment in inducing the NF-κB/NLRP3 signaling pathway and the gut-brain axis in the therapy of spasm after stroke. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Electric Acupuncture Treatment Promotes Angiogenesis in Rats with Middle Cerebral Artery Occlusion Through EphB4/EphrinB2 Mediated Src/PI3K Signal Pathway.

BACKGROUND: Cerebral infarction is one of the most common causes of disability and death worldwide. It is reported that electric acupuncture was able to improve the prognosis of cerebral infarction by promoting angiogenesis. However, the corresponding signal pathways of angiogenesis promotes by electric acupuncture treatment needs to be further explored. METHODS: MCAO rat was employed as the animal model, and clopidogrel hydrogen sulfate treatment was set as the positive control. Behaviors of rats, H&E staining, and TTC-staining was used to evaluate the recovery of infarcted brain tissue and nervous function. After that, immunocytochemical and immunofluorescence staining was used to quantify the angiogenesis and compensatory circulation, which including the analysis of microvessel density, field/ microvessel area ratio, and microvessel diameter. Western blot and RT-PCR for the detection of the related signal molecule, PI3K, Src, and EphB4/ephrinB2. RESULTS: The neurologic impairment scores were decreased, and the brain tissue damage that showed with H&E and TTC-staining was relieved by the treatment of electric acupuncture in MCAO rat. The quantification of microvessel density and field/ microvessel area ratio was improved obviously, and the microvessel diameter was decreased which represent the angiogenesis of capillary in day 3 and 7 by the electric acupuncture treatment. We also found that the level of Src and PI3K was increased markedly followed by the up-regulation of EphB4 and EphrinB2 mRNA during the electric acupuncture treatment, and the pre-treatment of Src and/or PI3K inhibitor was able to disturb the angiogenesis of capillary. CONCLUSIONS: We proved that electric acupuncture was able to accelerate the recovery of infarcted brain tissue and nervous function in MCAO rat by the promotion of angiogenesis, which was regulated by EphB4/EphrinB2 mediated Src/PI3K signal pathway. Our study provides a potential therapy and theoretical basis for the clinical treatment of cerebral infarction by the use of electric acupuncture.