| ObjectiveBy invstergation the dynamic changes of inflammatory factors surrounding hematoma after Cerebral hemorrhage and its impact of electro-acupuncture therapy at different time points, comprehend the mechanism of inflammatory factors atfter ICH, study the possible intervention mechanism about inflammatory immune response of the electroacupuncture treatment after acute cerebral hemorrhage.MethodsChoosing120male Wistar rats were randomly divided into normal group, sham-operated group, model group and EA group, normal group, sham-operated group, with18rats each team, the model groups were divided into6h,24h,48h,72h,8days5points, with18rats each team, EA groups were divided into, with18rats each team. the model groups and EA groups Use the0.5u collagenase VII injected rats caudate nucleus to make the modelof experimental, we applied the baihui、shuigou、zusanli (both sides) for the EA group, In the corresponding time points after modeling6h,24h,48h,72h,the EA begined to Scalp-acupuncture therapy (Continuous-wave frequency of120beats/min, intensity of1mA, the treatment time30min, Once a day, five days in a row.) there wasn’t any treatment of other groups.In the corresponding time points rats were killed to detect of relevant indicators, Including to measure the brain water content by dry-wet weigh method and observe the morphological changes of perihematomaby He staining, The others were to measure the protein expressions of I CAM、MMP-2、MMP-9、 TIMP-1by Immunohistochemical and Western blotResult(1)The results of HE staining indicate:There were no significant swelling of normal group and sham operation group of nerve cells, arranging in neat rows, structural integrity and a clear nucleolus. The model group6h rat caudate nucleus hematoma can be seen large areas of hemorrhage and spotting red blood cell infiltration, necrosis of nerve cells, the medulla visible interstitial part of the sheet with mild edema;24h can be seen a large number of vacuoles, neutrophil and lymphocyte infiltration, inflammation obvious; serious48h hematoma surrounding brain edema, seen a large number of microglia infiltration and inflammatory cell infiltration;72h brain tissue within the brain tissueedema and cell gap cavitation is most evident around can be seen more and inflammatory cell infiltration; eight days of the model group with mild edema around the hematoma;6h EA group of brain tissue seen in spotting, stromal cells, any mild edema, visible part of the neutrophil infiltration and glial cells; with the model group6h is quite similar. Visible edema within the brain tissue of24h EA group significantly reduced necrosis significantly reduced, very small amount of bleeding, certain degree of neutrophil infiltration. 48h EA group, the edema was reduced, visible see a small amount of neutrophil infiltration,72h EA group visible edema disappeared, with the model group, a marked improvement in eight days. EA treatment can significantly reduce inflammation, reduce the infiltration of inflammatory cells.(2) The water content of brain of Cerebral hemorrhage rats at each time point tissue indicate:6h model group, brain water content increase,24h, the bleeding side of the brain water content was significantly increased, peaked at72h,24-72h to maintain a high level and then decreased, a significant difference (P<0.01) compared with normalthe corresponding point in time the EA group compared with model group, EA group6h6h model group no significant difference between the rest of the EA of the corresponding point in time with the model group and there was significant sex difference (P<0.01), compared with eight days of model group was statistically significant (P<0.05),6h,24h,72h, the EA group after electro-acupuncture treatment can significantly reduce the water content of brain tissue among the groups no significant sexdifferences.(3) ICAM-1immunohistochemistry and Western blot results show: Model groups perihematoma have varying degrees of ICAM-1positive endothelial cells and neurons,6hours after intracerebral hemorrhage around blood vessels and nerve cells are the expression of ICAM-1,24h,48h, ICAM-1expression continued to increase,72hreached the peak at a higher level of expression in the8d any. EA group compared with the corresponding time point of the model group, ICAM-1expression, a significant sex difference (P<0.01),6h group of eight days of the EA group was not statistically significant (P>0.05), EA24h group,48h,72h group and8days group significantly with sex differences (P<0.05), while between EA24h group,48h,72h group group comparison no obvious significant difference (P>0.05), electro-acupuncture can reduce ICAM1expression(4) MMP-9、TIMP-limmunohistochemistry and Western blot results show:Normal group, no significant MMP-9, TIMP-1expression, the model control group6h visible significant expression of MMP-9,24h expression gradually increased and reached a peak at48h group, a small amount of low level of expression at day8; model controlgroup of TIMP-1expression and MMP-9expression of each group. After electro-acupuncture treatment in addition to the6h group EA, the EA treatment group time points of MMP-9expression compared with the model control group at each time point comparison has significantly reduced; In addition to the6h group of electro-acupuncture, electro-acupuncture treatment group a TIMP-1expression, the point in time relatively significant increase compared with the model control group at each time point. Statistical comparison:6h group of electro-acupuncture treatment control group6h group and8days respectively, and the model comparisons are not statistically significant (P>0.05), while the rest of the EA group and the corresponding time point of the model control group, MMP-9of TIMP-1of expression exist with significant differences (P<0.01), EA24h group,48h,72h group and8days group has significantly with the nature of the differences (<0.05), electro-acupuncture can reduce the cerebral hemorrhage around the hematoma in rats, MMP-9and increased TIMP-1expression.(5) MMP2immunohistochemistry and Western blot results show: Normal group and sham operation group, the expression of MMP-2was no significant difference (P>0.05), each model group compared with the sham group and normal group has a significant sex difference (P<0.01); model control group6h visible MMP-2a significant expression to24h continued to increase to the highest peak,48h-72h gradually reduced, but still significantly higher than the normal group, eight days, there is a high level of expression; after electro-acupuncture therapy, except for electro-acupuncture treatment group6h, and the remaining EAsignificantly reduce the treatment group at each time point MMP2in6h group of electro-acupuncture treatment6h group and8days compared with the model control, MMP-2expression were not statistically significant (P>0.05); the rest of the EA treatment groupcompared with the model control group the corresponding time points of MMP-2expression of differences (P<0.01), compared with eight days of model control group, significant sex differences (P<0.01). EA can reduce intracerebral hemorrhage in rats hematoma surrounding tissue expression of MMP-2.Conclution:The study shows that after the acute phase of cerebral hemorrhage, electro-acupuncture treatment has significantly reduce ICAM-1, MMP-9, MMP-2expression and increased TIMP-1expression,So that induced anti-inflammatory response after anti-cerebral hemorrhage, which is the result of acupuncture’s multi-target, multi-channel role... |
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