Neuroprotective Effect Of Electroacupuncture Exerts In Ischemic Stroke And The Effect On Neural Stem/Progenitot Cells Via Shh Signal Pathway

Objective:through the electroacupuncture of Quchi"LI11" and Zusanli"ST36" at the affected side limbs in the left side Middle Cerebral Artery ischemia rats, and the observation of the the expression of sonic hedgehog signaling pathways related molecule and the changes of Nestin in the left cortex after the ischemia, to discuss the mechanisms of" LI11" and " ST36" in the treatment on cerebral ischemia.Methods:72SD rats were randomly divided into24case of Sham-operation group and48case of MCAO group, the left Middle Cerebral Artery Occlusion (MCAO) reperfusion method was used on MCAO group for the Ischemia Reperfusion Injured rat molding. Graded neurologic behavioral scores in rats at2h after reperfusion, eliminated the unqualified rats, and rats of the MCAO group were randomly divided into Model group and the Electroacupuntrue group, in which were treated daily electroacupuncture intervention, the acupuncture points were selected at "LI11" and "ST36" in the affected side limbs of the rats, within1-20hz frequency,1times per day,30min/times, the total treatment was of3days. The neurologic behavioral scores were tested on the model groups and the electroacupuncture group respectively after24h,48h and72h after ischemia reperfusion.The rats were sacrificed at72h after ischemia reperfusion, TTC staining method was used to observation the infarction volume changes of the MCAO rats after electroacupuncture treatment, the neurologic behavioral scores were taken to evaluate nerve functional improvement of MCAO rats, animal behaviour changes of rats were detected by Homecage Scan real-time detection. Western blot and realtime PCR were taken to detect the protein and gene expression of Shh signaling pathways related factor, the morphology changes of nestin, GFAP of each group was detected by immunohistochemistry and immunofluorescence.Results:(1)24h after the ischemia reperfusion, there were no statistically significant differences between electroacupuncture group and model group in the neurobehavioral scores (P>0.05),72h after the reperfusion, the nerve function defect symptoms of electroacupuncture group were improved significantly than that in the model group (P<0.05);(2)Homecage Scan displayed the distance traveled in the cages of the model group were shorter than Sham-operation group (P<0.05), while the electroacupuncture group was longer than model group (P<0.05), grooming time and activity of turning of the model group were shorter than Sham-operation group (P<0.05), while the electroacupuncture group were longer than in model group (P<0.05);(3)And the cerebral infarction volumes of electroacupuncture group were also significantly reduced contrasted with the model group (P <0.05);(4) the protein and gene expression of Shh signaling pathways related factor in the cortex of the model group and the electroacupuncture group were increased contrasted with the sham-operation group, and the electroacupuncture group were increased more significantly than in the model group (P<0.05);(5) Immunohistochemistry revealed nestin, GFAP expression in the ischemia side cortex were significantly higher than in the sham-operation group (P<0.05), while the electroacupuncture group was increased significantly than in the model group (P<0.05). Immunofluorescence showed nestin/GFAP positive expression appeared after cerebral ischemia, the expression of electroacupuncture group was more than that of the model group.Conclusion:Shh pathway activation may participate in the neuroprotection of electroacupuncture at "LI11" and "ST36" in ischemia stroke.