| Purpose:To explore the angiogenic mechanism of eye-acupuncture treating cerebral ischemia reperfusion through established the cerebral ischemiareperfusion injury model in rats, and Immunohistochemisty SABC method was used to investigate the expression of Brain-Derived Neurotrophic Factor (BDNF),tyrosine kinase B(TrkB) and P75 neurotrophin receptor(P75NTR) in the brain tissue of the rat.Material and method:40 SPF healthy male Wistar rats (Weight:300±20g) were randomly divided into 4 groups in 8 cages:shame-operated group, model group, body-acupuncture group and eye-acupuncture group. Each group has 10 rats. There were 5 rats in each cage. After adaptive feeding for one week, middle cerebral artery occlusion model established by improved modified suture method, in accordance with the Zea-Longa signs of neurological defect standards assessed(NSS) after cerebral ischemia reperfusion (CIRI); There were 0-4 five grades. The model group,body-acupuncture group and eye-acupuncture group select the rats in level 1,2 and 3。After CIRI, the sham-operated group and model group did not take any treatment, eye-acupuncture group take the eye-acupuncture treatment in accordance with human eyes on the focal area, take Shangiiao area, Xiajiao area, liver area and kidney area. Acupunctured for 2 times which was in 20 minutes per time and every 12 hours during animal survival. The body-acupuncture group acupunctured the Quchi and Zusanli acupoint used as the same times as eye-acupuncture group. Specimen collection:Successfully reproduced in the model after 24 hours, in accordance with 0.35mL/100g Urethane anaesthetized, rapid perfusion of the ascending aorta, take the hippocampus tissue reserved。Using Tripenylteazolium chloride (TTC) dyeing observed for ischemic infarction and judged whether to establish MCAO model success. Immunohistochemisty SABC method was used to investigate the expression of BDNF,TrkB and P75NTR and with SPSS15.0 software statistics using single factor analysis of variance among groups and using paired test to analyze the different NSS before and after treatment in each group.Results:1. The situation of CIRI model establishedThere is no rat dead during the experiment. In accordance with the Zea-Longa signs of neurological defect assessed NSS after cerebral ischemia reperfusion showed:①There was no signs of neurological defect in the sham-operated group and NSS was 0 score.②Compared with the sham-operated group, there had the signs of neurological defect in the other three groups without statistically significant differences among three groups. These displayed the MCAO model wassuccesses;After Tripenylteazolium chloride (TTC) dyeing observed that the normal brain tissues were red and ischemia tissues were white which improved to establish MCAO model success.2. Effect of acupuncture on NSS score of the rat after cerebral ischemia reperfusionAfter CIRI model were established for 24 hours:①in model group, the signs of neurological defect had no obvious improvement, and there was no change in NSS score.②Through paired test, compared with the pre-treatment score, the signs of neurological defect assessed decreased (P<0.05)and the symptoms alleviated after treatment in body acupuncture group which showed that body-acupuncture treatment was effective.;③Through paired test, compared with the pre-treatment score, the signs of neurological defect assessed decreased (P<0.01)and the symptoms alleviated after treatment in eye acupuncture group which showed that eye-acupuncture treatment was effective; Compared with those in the model group, the signs of neurological defect decreased and the ssymptoms alleviated in the body acupuncture group and eye acupuncture group after the acupuncture, which revealed the acupuncture can improved the signs of neurological defect.3. Effect of eye-acupuncture on expression of the BDNF in the hippocampus tissue of the rat after cerebral ischemia reperfusionCompared with those in the sham-operated group, the expressions of BDNF in the other three groups were significantly increased (P<0.01); Compared with those in the model group, the expressions increased in the body-acupuncture group (P<0.05) and had Significant difference in the eye-acupuncture group (P<0.01); Compared with those in the body-acupuncture group, the expressions of BDNF increased (P<0.05) in the eye-acupuncture group。Above all, the expressions of BDNF was the strongest in eye-acupuncture group.4. Effect of eye-acupuncture on expression of the TrkB in the hippocampus tissue of the rat after cerebral ischemia reperfusion.Compared with those in the sham-operated group, the expressions of TrkB in the model groups were increased (P<0.05); the expressions significant increased in the body-acupuncture group and the eye-acupuncture group (P<0.01); Compared with the model group, the expressions of TrkB increased in the body-acupuncture group (P<0.05) and the expressions significant increased in the eye-acupuncture group (P<0.01); Compared with those in the body-acupuncture group, in the eye-acupuncture group the expressions of TrkB significantly increased (P<0.01)。Above all, the expressions of TrkB were the strongest in eye-acupuncture group.5. Effect of eye-acupuncture on expression of the P75NTR in the brain tissue of the rat after cerebral ischemia reperfusion。Compared with those in the sham-operated group, the expressions of P75NTR in the model group and body-acupuncture group were significantly increased (P<0.01), in the eye-acupuncture group the expressions of P75NTR increased (P<0.05) Compared with those in the model group, the expressions decreased in the body-acupuncture group (P<0.05) and decreased significantly in the eye-acupuncture group(P<0.01); Compared with those in the body-acupuncture group, the expressions decreased obviously in the eye-acupuncture group (P<0.01)。Above all, the expression of P75NTR was the strongest in model group, and decreased rapidly in the eye-acupuncture group.Conclusion:1. After the acupuncture, the signs of neurological defect alleviated in the cerebral ischemia reperfusion model rats。2. Acupuncture improving ischemic stroke through promoting the expression of BDNF and TrkB in the hippocampus tissue, the effect of eye-acupuncture treatment is better than body-acupuncture treatment in CIRI early stage.3. Acupuncture improving ischemic stroke through restraining the expression of P75NTR in the hippocampus tissue, the effect of eye-acupuncture treatment is better than body-acupuncture treatment in CIRI early stage. |
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