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Pre-electroacupuncture Ameliorates Cerebral Ischemia-reperfusion Injury by Inhibiting Microglial RhoA/pyrin/GSDMD
Hao Fang1,2,3, Ling-Ling Fan2,3, Ye-Ling Ding2,3
1State Key Laboratory of Traditional Chinese Medicine Syndrome, The Second Affiliated Hospital of Guangzhou, University of Chinese Medicine, Guangzhou, 510405, China.
Abstract:
Cerebral ischemia reperfusion injury is a severe neurological impairment that occurs after blood flow reconstruction in stroke, and microglia cell pyroptosis is one of its important mechanisms. Electroacupuncture has been shown to be effective in mitigating and alleviating cerebral ischemia reperfusion injury by inhibiting neuroinflammation, reducing cellular pyroptosis, and improving neurological function. In this experiment, we divided the rats into three groups, including the sham operation (Sham) group, the middle cerebral artery occlusion/reperfusion (MCAO/R) group, and the pre-electroacupuncture (EAC) group. Pre-electroacupuncture group was stimulated with electroacupuncture of a certain intensity on the Baihui (GV 20) and Dazhui (GV 14) of the rat once a day from the 7th day to the 1st day before the MCAO/R operation. The extent of cerebral infarction was detected by TTC staining. A modified Zea-Longa five-point scale scoring system was used to determine neurologic function in MCAO rats. The number of neurons and morphological changes were accessed by Nissl staining and HE staining. The cellular damage was detected by TUNEL staining. In addition, the expression levels of RhoA, pyrin, GSDMD, Caspase1, cleaved-Caspase1, Iba-1, CD206, and ROCK2 were examined by western blotting and immunofluorescence. The results found that pre-electroacupuncture significantly attenuated neurological impairment and cerebral infarction compared to the post-MCAO/R rats. In addition, pre-electroacupuncture therapy promoted polarization of microglia to the neuroprotective (M2) phenotype. In addition, pre-electroacupuncture inhibited microglia pyroptosis by inhibiting RhoA/pyrin/GSDMD signaling pathway, thereby reducing neuronal injury and increasing neuronal survival in the MCAO/R rats. Taken together, these results demonstrated that pre-acupuncture could attenuate cerebral ischemia-reperfusion injury by inhibiting microglial pyroptosis. Therefore, pre-electroacupuncture might be a potential preventive strategy for ischemic stroke patients.
Insights
Pre-electroacupuncture pretreatment effectively reduced brain damage and neurological deficits after stroke in rats. It inhibited microglial pyroptosis and promoted neuroprotective M2 microglia polarization, suggesting a potential preventive stroke therapy.
Area of Science:
- Neuroscience
- Neurology
- Cellular Biology
Background:
- Cerebral ischemia reperfusion injury is a major cause of stroke-related disability.
- Microglia cell pyroptosis plays a critical role in the pathogenesis of cerebral ischemia reperfusion injury.
- Electroacupuncture (EAC) shows promise in mitigating neuroinflammation and improving outcomes.
Purpose of the Study:
- To investigate the protective effects of pre-electroacupuncture (pre-EAC) on cerebral ischemia reperfusion injury (MCAO/R) in rats.
- To elucidate the underlying mechanisms involving microglia pyroptosis and polarization.
Main Methods:
- Rats were divided into sham, MCAO/R, and pre-EAC groups.
- Pre-EAC was administered daily for 7 days prior to MCAO/R induction.
- Cerebral infarction, neurological function, neuronal damage, and molecular markers (RhoA, pyrin, GSDMD, Caspase1, Iba-1, CD206, ROCK2) were assessed using TTC staining, neurological scoring, Nissl/HE staining, TUNEL assay, Western blotting, and immunofluorescence.
Main Results:
- Pre-EAC significantly reduced cerebral infarction extent and neurological impairment compared to the MCAO/R group.
- Pre-EAC promoted microglia polarization towards the M2 (neuroprotective) phenotype.
- Pre-EAC inhibited microglia pyroptosis by suppressing the RhoA/pyrin/GSDMD signaling pathway, reducing neuronal injury and enhancing neuronal survival.
Conclusions:
- Pre-electroacupuncture effectively attenuates cerebral ischemia-reperfusion injury in a rat model.
- The protective effects are mediated by the inhibition of microglial pyroptosis and modulation of microglial polarization.
- Pre-EAC represents a potential preventive therapeutic strategy for ischemic stroke.
