| Objective:To observe the opening effect of normal control and Ischemia-Reperfusion in recovery period Rats’ Blood-Brain Barrier with EA treatment under different EA time courses,and make clear this effect does not cause brain edema.In the meantime,screen out the optimal time course of EA treatment.Besides,demonstrating that this method can induce NGF to penetrate the Blood-Brain Barrier.Methods:Part one:Influence of EA under different time courses on the penetration amounts of Evans blue of rats,including Three experiments.Experiment 1:Influence of EA under different time courses on the penetration amounts of Evans blue of normal rats.The male SD rats were randomly divided into the normal control group,EA-1 min、5 min、15 min、30 min、45 min and 60 min group,there are 7 groups(16 rats each group).The rats in each group were randomly then divided into 2 subgroups(n=8),8 of them for the brain water content determination,the other 8 were for detecting the penetration amounts of Evans blue.EA groups used acupuncture needles of 0.30mm × 25mm,piercing GV20、GV26 acupoints,connecting the HANS acupoint nerve stimulator,current 2mA,frequency 100Hz,operating the corresponding long time of EA intervention.One minute before the ending of EA intervention,rats were injected 2%EB saline by cauda vein using indwelling needle(4mL/kg).The rats injected with anesthetics 30min after intervention.Normal control group without treatment of EA,rats were injected 2%EB saline by cauda vein,after 30min anesthesia is executed.The brain water content was measured by drying wet method.The penetration amounts of Evans blue was detected by fluorescence method.Experiment 2:Influence of EA under different time courses on the penetration amounts of Evans blue of MCAO/R rats in recovery period.Establishing the unilatera MCAO/R model SD rats using Zea Longa modified thread occlusion method(Surface cerebral blood flow was measured by laser Doppler flowmetry),then carrying out neural behavioral score after the rats wake up.The rats were randomly divided into the model control group,EA-1 min、5 min、15 min、30 min、45 min and 60 min group,when 21 days after operation(integrity of the blood-brain barrier in rats),setting up another sham-operated group(except for ligating artery and inserting intraluminal monofilament,the rest of procedure is same as the other groups),there are 8 groups(16 rats each group).the rats in each group were randomly then divided into 2 subgroups(n=8).The same detection methods like Experiment 1.Experiment 3:Further optimized of time courses of EA on the penetration amounts of Evans blue of MCAO/R rats in recovery period.Based on the analysis of the results of experiment 1 and experiment 2,the electroacupuncture time courses were further optimized.The same mode-1 method like Experiment 2.The rats were randomly divided into the modelcontrol group,EA-4 min、6 min、8 min、12 min、15 min and 45 min.when 21 days after operation.Setting up another sham-operated group,there are 9 groups(16 rats each group).the rats in each group were randomly then divided into 2 subgroups(n=8),The same detection methods like Experiment 1.Part two:Influence of EA on the penetration amounts of NGF of MCAO/R rats.Based on the result of Experiment 1,choose 8 minutes as optimal EA time course,then 16 SD rats for Establishing the unilatera MCAO/R model,they were randomly divided into the model control group、EA-8 min group when 21 days after operation,setting up another normal control group,there are 3 groups(8 rats each group).EA groups used acupuncture needles of 0.30mm × 25mm,piercing GV20、GV26 acupoints,connecting the HANS acupoint nerve stimulator,current 2mA,frequency 100Hz,8 min.One minute before the ending of EA intervention,rats were injected NGF by cauda vein using indwelling needle(10 ug/kg).The rats injected with anesthetics 30min after intervention.Normal control and model group without treatment of EA,rats were injected NGF by cauda vein,after 30min anesthesia is executed.Perfusion and fetched out the cerebral cortex,the content of NGF in brain tissu-eswas measured by ELISA testing.Result:Part one:Experiment 1.(1)The brain water content in each group was not different(P>0.05).(2)The penetration amounts of Evans blue in EA-1 min group、EA-5 min group、EA-15 min group were apparently higher than normal control group(P<0.01).The penetration amounts of Evans blue in EA-5 min group was higerer than other groups(P<0.01).Experiment 2.(1)The brain water content in each group wa-s not different(P>0.05).(2)EA-5 min g-roup、EA-15 min group、EA-30 min group were apparently higher than model contro-1 group and sham-operated group(P<0.01).The penetration amounts of Evans blue in EA-5 min group was higerer than other groups(P<0.01).In addition to the EA-5 min group,the penetration amounts of Evans blue in EA-15 min group was higerer than other groups(P<0.01).Experiment 3.(1)The brain water content in each group was not different(P>0.05).(2)EA-4 min group、EA-6 min group、EA-8 min group、EA-12 min group were appare-ntly higher than model control group and sham-operated group(P<0.01).In addition to the EA-8 min group,the penetration amounts of Evans blue in EA-6 min group was higerer than other groups(P<0.01).EA-6 min group was lower than EA-8 min group(P<0.05).The penetration amounts of Evans blue in EA-8 min group was higerer than other groups(P<0.01).Part two:The content of NGF increased significantly in EA group compared with normal control group.and there is a significant difference between them(P<0.01).the content of NGF in EA group was apparently higher than model control group(P<0.01).Conclusion:1.EA treatment at GV20 GV26 can induce EB to penetrate the normal control and Ischemia-Reperfusion Rats’ Blood-Brain Barrier in varying degrees,and this effect does not cause brain edema.2.Furthermore,the effect of EA on the opening of BBB is different,there was difference between different EA time courses for opening the BBB,among them,EA-8 min was the best time;3.EA treatment at GV20 GV26 under 8 minutes cansignificantly increase the content of NGF i-n brain;3.E A-8 min at GV20 GV26 of MCAO/R rats,also has a good effect on the penetration amounts of NGF. |
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