| Objective: To explore the effects of electro-acupuncture on the expression of growth associated protein (GAP-43) in hippocampus, striatum and motor cortex of neonatal asphyxial cerebral palsy rats, and to explore the significance and mechanism of electro-acupuncture in treatment of cerebral palsy.Methods: 1) 72 7d Sprague-Dawley (SD) rats were randomizd into 3 groups, sham operation group(n=24),CP model group(n=24) and CP+ electro-acupuncture group (n=24). Each group were randomizd into 4 time point groups (n=6 in each group); 2) Cerebral palsy model was established by occlusion of the left common carotid artery and inhalation of 8% oxygen plus 92% nitrogen 2 hours. The rat which was found turnover unability, spontaneously turn left round and balance disorder was regarded as success model, and were randomizd into CP model group and CP+ electro-acupuncture group. sham operation group jiust separat the left common carotid artery, three groups were feeded by maternal rats after operation; 3)The electro-acupuncture group began treatment in 24h after operation. Acupuncture at Baihui, Dazhui, Jizhong, Houhui point and connect a G6805-II Electroacupuncture Therapeutic Apparatus, twice a day, the rats in each time point group were sacrificed in the last day 2h after second treatment; 4) The section of brain anterior 1～3mm from bregma (to observe motor cortex and striatum) and posterior 2～3mm from bregma (to observe hippocampus) was sectioned serially coronoidly, 4μm in thickness, by Immunohistochemical Staining (SABC mothod) and HE staining; 5) Using Leica IM50 image collection system,6 integrity and no overlap image with high power light microscope were selected from hippocampus, striatum and motor cortex in different section. And Detecting the mean optical density and surface density of GAP-43 in every image by Image-Pro Plus, to observe the diversity expression of GAP-43 in brain.Results: 1)From light microscopic observations, HE staining showed that sham operation group cerebral morphology were normal in each time point, CP model group and CP+ electro-acupuncture group showed that some cell presented some morphologic features of apoptosis in different time point and different field, including cell shrinkage, plasma membrane blebbing, nuclear condensation and formation of apoptotic bodies. Non-apoptotic Nerve cells presenting mainly as with raritas plasma, light staining and interstitial edema, there was a decreasing trend of apoptotic cells in 7d, 14d, 21d rats'brain, neuronal degeneration gradually disappeared; 2)Each group GAP-43 immunohistochemical staining positive staining matter appeared brown, located in neural soma and axon. Sham operation group immunoreactive cell with light staining and small amount. And CP model group and CP+ electro-acupuncture group a large number of GAP-43 positive pyramidal layer cells with brown-yellow granulations in endochylema was found in the in hippocampus, striatum and motor cortex; 3) Compared with sham operation group, expression of GAP-43 average optical density and positive area rates were more higher in CP model group and CP+ electro-acupuncture group (P<0.01) . The average optical density and positive area rates of GAP-43 in CP+ electro-acupuncture group was significantly higher than those of CP model group(P<0.05或P<0.01). Expression of GAP-43 was found at each time point in the sham operation group, but the expression was no difference. The expression peak of GAP-43 in CP model group and CP+ electro-acupuncture group was on 7d, and than declined.Conclusions: Electro-acupuncture could promote expression of GAP-43 in neonatal asphyxial cerebral palsy rats, accordingly could promote nerve cell regeneration and synapse reconstitution, which may contribute to the effect of acupuncture in improving cerebral palsy. |
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