Study Of Time-effect Relationship With Electro-acupunture In Treating Acute Cerebral Hemorrhage

Objective:We intend to present the main issues isⅦcollagenase-induced intracerebral hemorrhage rat model,the use of morphological and biochemical techniques,in the point of the inflammatory immune response after experimental intracerebral hemorrhag,observe and study of effect for different time point TNF-α,MMP-9's expressi on,explore the different times of electro-acupuncture in treating acute cerebral hemorrhage time-effect relationship,determine the best time to intervene for the clinical treatment to provide a solid experimental basis.Methods:Choose healthy wistar male rats 56,weight at 200-250g. According to the weight numbers into the random number table is divided into normal group,model group,3h and 72h Electroacupunc ture group,model group is divided into model 1,model 2,model 3, model 4 group;every group rat have 8 rats.Model used 0.5uⅦcollagenase injection rat caudate putamen(the former after fontanelle 1mm,median sagittal suture to the right 4mm,deep 5mm)-induced rat brain hemorrhage.Electro-acupuncture group of cerebral hemorrhage in rats using Shuigou points,Neiguan(bilateral),Sanyinjiao points (bilateral).Respectively,3h and 72h electro-acupuncture group when the Success of model after 3h,72h begin acupuncture,Shuigou and right ears of rats received a pair of electrodes,Sanyinjiao ipsilateral Neiguan received a pair of electrodes,continuous wave, frequency 120 times / min,intensity of 1mh,Every time the treatment last 30min,once daily acupuncture,acupuncture 5 times.Model 1 and Model 2 separately whenthe success of model after 3h,72h instantly killed,model 3 group and model 4 group separately when the success of model after the +5 d executed derived.The seven rats were used 10%chloral hydrate by intraperitoneal injection of anesthesia,thoracotomy via the ascending aorta followed by filled with normal saline 100ml,4℃Precooling containing 4%paraformaldehyde in 0.01mol / L phosphate buffer(PBS, pH7.4) 400ml.Completed irrigation and brain tissue was collected, placed in 4%formaldehyde solution after overnight fixation. Injection point as the center coronal paraffin section,film thickness 5μm,cut 10,affixed to a piece of Anti-APES glass baking for stained.HE staining,immunohistoch emical detection of tumor necrosis factor-α(Tumor necrosis factor-α,TNF-α) and matrix metalloproteinase -9(matrix metallo proteinase-9,MMP-9),with high-power microscope at 400 times,randomly checked the five non-overlapping field of vision around the hematoma,counting TNF-αand MMP-9 positive cells,statistical analysis.Results:(1) HE staining Results:In normal brain tissue no obvious abnormalities;model of brain tissue within one group see a large number of cerebral hemorrhage,neurofibrillary tangles,fracture, there was no obvious microglial cell infiltration in brain tissue; Model group 2 see a small amount of bleeding,some microglia cell infiltration(some cells within a bubble) and substantial liquefaction necrosis(soft bad cribriform foci);model group 3 shows that a large number of brain microglia cell infiltration and necrosis witha small amount of bleeding;model group 4 liquefied brain lesions can be seen to narrow and substantial microglial cell infiltration;3h electro-acupuncture group brain tissue seen in foam cell infiltration and hemorrhage;72h electro-acupuncture group brain tissue can be seen within necrotic foci markedly reduced microglial cells infiltration,a very small amount of bleeding.(2) Immunohistochemical results:①The brain tissue TNF-α' s expression:in model group,the expression of TNF-αwere higher than the normal group(P＜0.01), a very significant difference;model 2 group' s expression was higher than the model 1 and model 4 group(P＜0.01),a very significant difference;the positive expression of 3h electro-acupuncture group was lower than the model 2 group(P＜0.01), a very significant difference;the positive expression of 3h electro-acupuncture group was lower than the model 3 group(P＜0.05), a significant difference;the positive expression of 72h electro-acupuncture group was lower than the model 2 group(P＜0.01), a very significant difference;the positive expression of 72h electro-acupuncture group was lower than 3h electro-acupuncture group(P＜0.05),a significant differences.②The brain tissue MMP-9' s expression:model 1 group' s expression of MMP-9 was higher than normal(P＜0.05),a significantly different;model 2,model 3,model 4 group,the expression of MMP-9 were higher than normal group(P＜0.01),a very significant difference;model 2 group' s expression was higher than the model 1 and model 4 group(P＜0.01),a very significant difference;the positive expression of 3h electro-acupuncture group was higher than the model 1(P＜0.05),a significantly different;the positive expression of 3h electro-acupuncture group was lower than the model,2 group(P＜0.01),a very significant difference;the positive expression of 3h electro-acupuncture group was lower than the model 3 group(P＜0.05 ),a significant differences;the positive expression of 72h electro-acupuncture group was lower than the model 2 group(P＜0.01),a very significant difference;the positive expression of 72h electro-acupuncture group wsa lower than the model 4 group(P＜0.05),a significant differences;the positive expression of 72h electro-acupuncture group was lower than 3h electro-acupuncture group(P＜0.05),a significant differences.Conclusion:(1) inflammatory immune response involved in the formation of brain edema around brain haematoma after intracerebral hemorrhage, probably an important factor of cerebral hemorrhage injury.(2) electro-acupuncture possible by reducing brain tissue TNF-αand MMP-9 levels after cerebral hemorrhage,inhibition of induced by their inflammatory response,reducing their damage on blood-brain barrier(blood brain barrier,BBB),there is a certain degree of repair on cerebral edema and injure after cerebral hemorrhage.Electroacupuncture Treatment of cerebral hemorrhage certainly has some effect.(3) The TNF-αand MMP-9' s positive expression of 72h electro- acupuncture group was lower than 3h electro-acupuncture group. Possibly,the therapeutic effect of 72h electro-acupuncture group was better than 3h electro-acupuncture group.