| Objective:Based on the theory of opposing needling,observe the role of acupuncture in improving the motor function of limbs of middle cerebral artery occlusion(MCAO)model rats,and preliminary explore its effect mechanism.Method:1.MCAO models were prepared by using different thread plugs(self-made paraffin thread plugs,silicone thread plugs)and different anesthetics(chloral hydrate,isoflurane),and the success rate of the model and the survival rate within 7 days were observed to select more suitable model preparation conditions.2.Based on the establishment of appropriate model preparation conditions,four acupoints of Hegu,Neiguan,Zusanli and Yanglingquan were selected to perform electroacupuncture treatment on the healthy,affected and bilateral points of MCAO model rats.Before the model preparation,24 hours after the model preparation and 7 days after the acupuncture,the rats were subjected to behavioral scoring,and the model rats were subjected to TTC staining and brain water content determination to observe their therapeutic effects.3.Western Blot technology was used to detect the expression of GABAAR?1 protein and GAP-43 protein in the bilateral cortex,and the high-throughput sequencing analysis of the healthy cortex of the model rats was used to initially explore the potential mechanism of giant spines to improve motor function after ischemic stroke.Results:1.Compared with the self-made paraffin thread plug group in the first study,the rats in the silicone thread suppository group had a higher modeling success rate and a higher 7-day survival rate(P <0.05);There was no difference in the success rate of the model(P> 0.05),but the 7-day survival rate was higher in the isoflurane group(P <0.05).2.In the efficacy observation study,the neurological deficit score and behavior score of each group of rats before the model preparation were 0;24 hours after the model preparation,except for the sham operation group,the model group,the healthy side acupuncture group,the affected side acupuncture group Compared with the model before preparation,the bilateralacupuncture group had significantly higher scores(P <0.05),and there was no statistical difference between the groups(P> 0.05),which was comparable.After 7 days of acupuncture,the scores of the sham-operation group remained unchanged,while the scores of the other four groups all decreased but were still higher than that of the sham-operation group(P <0.05).In comparison,the behavioral scores of each group were lower than that of the model group(P <0.05);compared between the groups of acupuncture,the scores of the bilateral acupuncture group were lower,but there was no statistical difference(P> 0.05).TTC staining showed that the other four groups of rats except the sham-operated group all had white cerebral infarction.The 24 h model preparation group had the largest cerebral infarction volume.After 7 days of acupuncture,the cerebral infarction volume of the four groups of rats decreased.Compared with the model group,the volume of cerebral infarction in the acupuncture group on the affected side and the bilateral acupuncture group was smaller(P<0.05).There was no statistical difference between the acupuncture groups(P> 0.05).The brain water content was highest at 24 hours after the preparation of the rat model.After 7 days of acupuncture,the brain water content decreased relatively.The model group,the healthy side acupuncture group,the affected side acupuncture group,the bilateral acupuncture group and the model 24 h after the preparation There were statistical differences between the groups(P <0.05).The water content of each acupuncture group was lower than that of the model group(P <0.05),but there was no statistical difference between the acupuncture groups.The above results suggest that the healthy side acupuncture group,the affected side acupuncture group,and the bilateral acupuncture group can reduce the behavior score,reduce the cerebral infarction volume,and reduce the brain water content,which are statistically different from the model group There was no statistical difference between the acupuncture groups.3.Changes in the expression of GABAAR?1 protein in the affected cortex: After 7 days of acupuncture intervention,compared with the sham operation group,the remaining groups all had a downward trend,but only the healthy side acupuncture group and the affected side acupuncture group had a statistically significant downward trend(P <0.05),there was no significant difference between the model group and the acupuncture groups(P> 0.05);the expression of GABAAR?1 protein in the healthy side cortex compared with the sham operation group,the other four groups showed a downward trend,but none Statisticaldifference(P> 0.05).Changes in GAP-43 protein expression in the affected cortex: After 7days of acupuncture intervention,the trend of the model group and the sham operation group decreased significantly,there was a statistical difference(P <0.05),compared with the model group The amount showed an upward trend,of which there was a statistical difference between the acupuncture group on the contralateral side and the acupuncture group on the affected side(P <0.05),and there was no statistical difference between the bilateral acupuncture group(P> 0.05);GAP-43 protein on the contralateral cortex The expression level of the model group showed a downward trend compared with the sham operation group,but there was no statistical difference(P> 0.05);the expression level of the acupuncture group compared with the model group showed an upward trend,but no statistical difference(P>0.05).4.The results of high-throughput sequencing analysis of the healthy side cortex showed that 11 genes with significant changes in expression were screened after the healthy side acupuncture.The enrichment of GO function was mainly in the change of cell composition and development process.After the acupuncture on the affected side,13 genes with significantly different expressions were screened out.The enrichment of GO function was mainly in the changes inside and outside the cell.Conclusion:1.Compared with the self-made paraffin wire plug(made based on the conditions of this laboratory),the purchased silicone wire plug with uniform specifications is more suitable for the preparation of MCAO mode.Compared with chloral hydrate anesthesia,isoflurane anesthesia is more suitable for the preparation of MCAO model.2.Acupuncture on the contralateral side,acupuncture on the affected side,and bilateral acupuncture can effectively improve the function of the MCAO model rats and reduce the area of cerebral infarction and the degree of cerebral edema.3.The improvement of the motor function of the affected limbs of MCAO model rats after 7days of acupuncture on the contralateral side and acupuncture on the affected side may be related to the increased expression of GAP-43 protein in the cortex of the affected side.4.The improvement of the motor function of the affected limb of MCAO model rats after 7days of acupuncture intervention may be related to the PI3K-Akt signaling pathway of thehealthy side cortex. |
PDF Full Text Request