| Objective:Observe the effects of acupuncture on neuroethology,cerebral infarction volume,blood-brain barrier(BBB)permeability,and cerebral water content after thrombolysis in rats with acute cerebral infarction,and clarify the effect of acupuncture on improving the safety of cerebral infarction thrombolysis;and explore its mechanism of action from the astrocyte approach.Methods:This experiment is divided into two parts.Experiment 1:Study on the effect of acupuncture on improving the safety of thrombolysis in cerebral infarction.In this study,a modified rat autologous thromboembolism method was used to prepare a cerebral infarction model,the "Xing Nao Kai Qiao" acupuncture method was used as the intervention method,and the tail vein injection of alteplase(rt-PA)was used as the thrombolytic method.The SD rats were randomly divided into sham operation group,model group,4.5h thrombolysis group,6h thrombolysis group,acupuncture+4.5h thrombolysis group,acupuncture+6h thrombolysis group,24 hours after modeling,the effects of acupuncture on neuroethology,cerebral infarction volume,BBB permeability,and brain water content of rats in each group were observed,and the effect of acupuncture on improving the safety of cerebral infarction thrombolysis was confirmed.Experiment 2:Study on the mechanism of acupuncture improving the safety of thrombolysis in cerebral infarction.Model preparation,intervention methods,and thrombolysis methods are the same as Experiment 1.On the basis of experiment 1,SD rats were randomly divided into sham operation group,model group,6h thrombolysis group,acupuncture+6h thrombolysis group,and the samples were taken 24h after modeling.Real-time PCR and Western Blot methods were used to detect the mRNA and protein expressions of astrocyte-related indexes(glial fibrillary acidic protein GFAP,aquaporin 4 AQP-4)in the cerebral cortex of rats in each group,to explore the mechanism of acupuncture improving the safety of thrombolysis in cerebral infarction.Results:1.Comparison of neurobehavioral scores of rats in each group:The model was scored 24h after modeling,and the neurobehavioral score of the model group was higher than that of the sham operation group(P<0.01).There was no significant difference in neurobehavioral scores between the 4.5h thrombolysis group and the acupuncture+4.5h thrombolysis group(P>0.05),and both were lower than the model group(both P<0.01).The neurobehavioral scores of acupuncture+6h thrombolysis group were lower than model group and 6h thrombolysis group(P<0.05,P<0.01).2.Comparison of cerebral infarction volume percentage of rats in each group:The percentage of cerebral infarction volume in the model group was higher than that in the sham operation group(P<0.01).There was no significant difference in the percentage of cerebral infarction volume between the 4.5h thrombolysis group and the acupuncture+4.5h thrombolysis group(P>0.05),and both were lower than the model group(both P<0.01).The percentage of cerebral infarction volume in the acupuncture+6h thrombolysis group was lower than that of the model group and the 6h thrombolysis group(both P<0.05).3.Comparison of BBB permeability in rats of each group:The EB content of the model group was higher than that of the sham operation group(P<0.01).There was no significant difference in EB content between the 4.5h thrombolysis group and the acupuncture+4.5h thrombolysis group(P>0.05),and both were lower than the model group(both P<0.01).The EB content of the 6h thrombolysis group was higher than that of the model group(P<0.01).The content of EB in the acupuncture+6h thrombolysis group was lower than that of the model group and 6h thrombolysis group(both P<0.01).4.Comparison of the percentage of brain water content of rats in each group:The percentage of brain water content in the model group was higher than that in the sham operation group(P<0.01).There was no significant difference in the percentage of brain water content between the 4.5h thrombolysis group and the acupuncture+4.5h thrombolysis group(P>0.05),and both were lower than the model group(both P<0.01).The percentage of brain water content in the 6h thrombolysis group was higher than that of the model group(P<0.01).The percentage of brain water content in the acupuncture+6h thrombolysis group was lower than that of the model group and the 6h thrombolysis group(P<0.05,P<0.01).5.Comparison of GFAP mRNA expression in the cerebral cortex of rats in each group:According to the results of Real-time PCR,the mRNA expression level of GFAP in the model group was higher than that in the sham operation group(P<0.01),The mRNA expression level of GFAP in the acupuncture+6h thrombolysis group was lower than that of the model group and 6h thrombolysis group(both P<0.01).6.Comparison of AQP-4 mRNA expression in the cerebral cortex of rats in each group:According to the results of Real-time PCR,the mRNA expression level of AQP-4 in the model group was higher than that in the sham operation group(P<0.01).The mRNA expression level of AQP-4 in the acupuncture+6h thrombolysis group was lower than that of the model group and the 6h thrombolysis group(both P<0.01).7.Comparison of GFAP protein expression in the cerebral cortex of rats in each group:According to the results of Western Blot,the protein expression level of GFAP in the model group was higher than that in the sham operation group(P<0.01).The protein expression level of GFAP in the acupuncture+6h thrombolysis group was lower than that of the model group and 6h thrombolysis group(P<0.05,P<0.01).8.Comparison of AQP-4 protein expression in the cerebral cortex of rats in each group:According to the results of Western Blot,the protein expression level of AQP-4 in the model group was higher than that in the sham operation group(P<0.05).The protein expression level of AQP-4 in acupuncture+6h thrombolysis group was lower than that of model group and 6h thrombolysis group(P<0.05,P<0.01).Conclusion:1.Simple thrombolytic therapy or acupuncture combined with thrombolytic therapy within the 4.5h time window of acute cerebral infarction have ideal brain protection effects and similar effects.2.Thrombolytic treatment of cerebral infarction outside the 4.5h time window is more likely to produce hemorrhagic transformation,cerebral edema and other side effects of thrombolysis.This study shows that after acupuncture in time,thrombolysis within6 hours of the onset of cerebral infarction can reduce the complications of thrombolysis and still have a safe effect.3.Acupuncture can down-regulate the mRNA and protein expressions of GFAP and AQP-4 in ischemic brain tissue,indicating that acupuncture can improve the thrombolytic safety by inhibiting the activation of astrocytes. |
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