"Baihui"(DU20)-penetrating"Qubin"(GB7)acupuncture on blood-brain barrier integrity in rat intracerebral hemorrhage models via the RhoA/ROCK Ⅱ/MLC 2 signaling pathway
摘要Background:Blocking the RhoA/ROCK Ⅱ/MLC 2(Ras homolog gene family member A/Rho kinase Ⅱ/myosin light chain 2)signaling pathway can initiate neuroprotective mechanisms against neurological diseases such as stroke,cerebral ischemia,and suba-rachnoid hemorrhage.Nevertheless,it is not clear whether and how disrupting the RhoA/ROCK Ⅱ/MLC 2 signaling pathway changes the pathogenic processes of the blood-brain barrier(BBB)after intracerebral hemorrhage(ICH).The present inves-tigation included the injection of rat caudal vein blood into the basal ganglia area to replicate the pathophysiological conditions caused by ICH.Methods:Scalp acupuncture(SA)therapy was performed on rats with ICH at the acu-puncture point"Baihui"-penetrating"Qubin,"and the ROCK selective inhibitor fasudil was used as a positive control to evaluate the inhibitory effect of acupuncture on the RhoA/ROCK Ⅱ/MLC 2 signaling pathway.Post-assessments included neurological deficits,brain edema,Evans blue extravasation,Western blot,quantitative polymer-ase chain reaction,and transmission electron microscope imaging.Results:We found that ROCK Ⅱ acts as a promoter of the RhoA/ROCK Ⅱ/MLC 2 signaling pathway,and its expression increased at 6h after ICH,peaked at 3 days,and then decreased at 7days after ICH,but was still higher than the pre-intervention level.According to some experimental results,although 3 days is the peak,7 days is the best time point for acupuncture treatment.Starting from 6h after ICH,the neurovascular structure and endothelial cell morphology around the hematoma began to change.Based on the changes in the promoter ROCK Ⅱ,a 7-day time point was selected as the breakthrough point for treating ICH model rats in the main experiment.The results of this experiment showed that both SA at"Baihui"-penetrating"Qubin"and treatment with fasudil could improve the ex-pression of endothelial-related proteins by inhibiting the RhoA/ROCK Ⅱ/MLC 2 signaling pathway and reduce neurological dysfunction,brain edema,and BBB per-meability in rats.Conclusion:This study found that these experimental data indicated that SA at"Baihui"-penetrating"Qubin"could preserve BBB integrity and neurological function recovery after ICH by inhibiting RhoA/ROCK Ⅱ/MLC 2 signaling pathway activation and by regulating endothelial cell-related proteins.
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