| Objective:To observe the Permeation Enhancing effect of the Rats’ Blood-Brain-Barrier with EA treatment at GV20/GV26 under Different Frequencies dilatational wave,and the impacts on theNGF in rat brain tissue,demonstrating that this method have the function of stimulating the BBB’s opening,meanwhile,screen out the optimal frequency of EA treatment.Method:Experiment 1:Establishing the unilatera MCAO/R model SD rats using Zea Longa modified thread-occlusion method(Surface cerebral blood flow(sCBF)was measured by laser Doppler flowmetry(LDF),Cerebral blood flow down to the basis of the value of 45 ±10%for the modeling standards,then carrying out neural behavioral score after the ischemia-reperfusion rats wake up).The rats were randomly divided into the model control group、2/15Hz group、2/30Hz group、2/50Hz group、2/100Hz group when 21 days after operation(integrity of theblood-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 6 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.1.The brain water content determination:Model control group and sham-operated group without treatment of EA,EA pretreatment each group used 0.30mm × 25mm,Hua Tuo brand acupuncture needles,piercing GV20、GV26acupoints,connecting the HANS acupoint nerve stimulator,current 2mA each 8min.the rats injected with anesthetics After intervention.Perfusion saline Through the left ventricle until when the fluid which flew out of the right auricle turned colourless.fetched out the cerebral cortex without olfactory and cerebellum,the brain water contentwas measured by drying wet method.2.detecting the penetration amounts of Evans blue:rats in each group were injected2%EB saline by caudal vein(4mL/kg),then electro-acupuncture is executed for 8 minutes,Among them,Model control group and sham-operated group without treatment of EA.After the intervention,the method of taking brain same as above,the penetration amounts of Evans blue was detected by fluorescence method.Experiment 2:based on the result of Experiment 1,24 SD rats were used for Establishing the unilatera MCAO/R model,they were randomly divided into the model control group、2/50Hz group,2/100 Hz group when 21 days after operation,setting up another normal control group,there are 4 groups(8 rats each group).The rats in all group were given an electro-acupuncture for 8 minutes after NGF injection,Model control group and normal control group without treatment of EA.the rats injected with anesthetics After intervention.Perfusion saline Through the left ventricle until whenthe fluid which flew out of the right auricle turned colourless.fetched out the cerebral cortex without olfactory and cerebellum,the content of NGF in brain tissueswas measured by ELISA testing.Result:1.The brain water content in each group was not different(P>0.05).2.In addition to the 2/30Hz group,the penetration amounts of Evans blue in other groups were apparently higher than Model control group and sham-operated group(P<0.05).there was no significance between model control group and sham-operated group(P>0.05).The penetration amounts of Evans blue in 2/50Hz group and 2/100Hz group all Increased compared with 2/15Hz group(P<0.05).The penetration amounts of Evans blue in 2/100Hz group was slightly lower than2/50Hzgroup,but was not significant(P>0.05).3.The content of NGF increased significantly in 2/50Hz group and 2/100Hz group compared with normal control group and there is a significant difference between them(P<0.01),and this phenomenon was most obvious in 2/100Hz group(P<0.001).there was no significance between model control group and normal control group(P>0.05).the content of NGF in 2/100Hz group was apparentlyhigher than Model control group(P<0.001).the content of NGF in normal control group,Model control group,and 2/50Hz group were apparently lower than Model control group(P<0.001).Conclusion:1.EB can be induced through the blood-brain barrier in rats and significantly increased by dilatational wave EA at Baihui(GV20),Shuigou(GV26).Among that,the dilatational waves with frequency 2/50Hz and 2/100Hz show the advantage,however they two have no difference from each other.In the meanwhile,different dilatational waves EA on promoting the open of Blood-Brain Barrier won’t cause cerebral edema.2.NGF can be induced through the blood-brain barrier in rats by dilatational wave 2/50Hz and 2/100Hz EA at Baihui(GV20),Shuigou(GV26).Among that,the dilatational wave with frequency 2/100Hz shows the advantage.3.Dilatational wave EA can induce NGF through the blood-brain barrier in rats,which provide an experimental foundation for the treatment of central nervous system diseases through the combination of acupuncture with medicine. |
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