| Objective:To investigate the effect of the brain tissue of the CP rat after conducting acupuncture and moxibustion treatment. Thus, provide the theoretical basis for the promotion of acupuncture and moxibustion treatment in clinical use for children with cerebral palsy.Methodology:The timing of intervention:1. Weigh after surgery (after birth 7d)and have a day rest.2. Have the over hanging test(OHT) and inclined plane test(IPT)after 2days of the surgery.3. Have the treatment 6 days on the 3rd-8th days of the surgery.4. Have the OHT and IPT test on the 9th day after the surgery and get rest.5. Have the treatment 6 days from thelOth to 15th day after the surgery,6. Have the OHT and IPT test on thel6th day,7. Have the Y maze test on the 16th-17th day,8. Weigh and be sacrificed on the 18th day (born 25d) Indicators observed:(a) Content of NE, DA,5-HT in monoamine neurotransmitters in brain tissues of CP(b) Content of excitatory amino acid (Glu) and inhibitory amino acid (GABA)in brain tissuesResults:1. Tests on Change of Body weight, Behavior, Ability of learning and memory(1)Weight:Multiple comparison results showed that:the normal group, sham operation operation group, moxibustion group, acupuncture moxibustion group were no significant difference between the four groups, model group and acupuncture group was no statistical difference neither. The first four groups compared with the other groups, P<0.01, both statistically significant weight gain can be considered the first four group weight gain more than two groups. Area group F=15.897, P> 0.05, shows that factors between area group was not statistically significant.(2)Behavioral test, Learning and memory tests(a) Behavioral testTreatment effect between groups statistical test results:OHT:F=2.34002, P<0.01; IPT:F=838.629, P<0.01. CP rats of different treatment methods on muscle strength, balance statistically significant differences in growth. The factors time and treatment interaction P<0.01, that the different treatment methods on muscle strength HIBD rats, balance role in promoting growth is different.Results of Between different groupsStrength (OHT):sham operation group) normal group〉acupuncture+moxibustion group〉moxibustion group〉model group〉acupuncture group. Balance (IPT): normal group〉sham operation group〉acupuncture+moxibustion group〉moxibustion group〉simple acupuncture group〉model group. Factors time and treatment interaction, the strength (OHT) growth trends: sham operation group〉normal group〉acupuncture+moxibustion group〉moxibustion group〉model group〉simple acupuncture group; Balance (IPT) growth trends:normal group〉sham operation group〉acupuncture+moxibustion group〉simple acupuncture group〉moxibustion group〉model group.(b) Learning and memory tests(Y-maze test)Results of Y-maze test show that:different treatment groups, F= 2.1793, P<0.01. Multiple comparison results showed that:normal group〉sham operation group〉acupuncture+moxibustion group〉moxibustion group〉simple acupuncture group〉model group.2. Pathological observation of brain tissue and cells(1) HE staining:normal light microscope under 10×20 times for each section were observed within 5 View pathological changes after cerebral ischemia. Each group were pathological changes in rat brain:brain tissue of paraffin, HE staining, nerve cells of pale red, blue-black nuclear was. Normal control group and the sham operation group structure of the normal cortical brain cells, clear nucleus, cytoplasm-rich, interstitial edema-free performance, hippoca(?)pal pyramidal cells in 2 to 3 layers, closely arranged, round and large nucleus, clear nucleolus. Model group, cortical thinning, part of the neuronal degeneration and smaller, the nucleus and cytoplasm of ambiguous nuclear fast shrunk triangles or irregular, nucleoli disappear. Reduce hippocampal pyramidal cell layer, arranged in sparse, irregular, smaller cell volume, nuclear condensation phenomenon can be seen. Acupuncture group, acupuncture combined with moxibustion moxibustion group and treatment group, there are different degrees of nerve cell degeneration, but significantly reduced the number of nerve cell degeneration, sporadic seen a number of injured cells, hippocampal pyramidal cell layer of cell morphology, arrangement normal.(2) Counting of experimental groups hippocampus nerve cell normal circumstancesCerebral cortex of rats in each group comparison of the number of normal nerve cells results show:the sham operation group and the normal control group (P> 0.05) difference was not significant. Sham operation group and model group (P<0.01) significant difference was significant. Acupuncture group, moxibustion group, acupuncture+moxibustion group compared with model group (P＜0.05) that the difference was statistically significant. And acupuncture group, moxibustion group, acupuncture+moxibustion group have a larger number of nerve cells than the model group. Acupuncture group, moxibustion group, acupuncture+moxibustion group comparison between the three differences (P> 0.05) were not statistically significant.3. UltrastructureNormal group:neuronal cell body formed integrity, the nuclei were round, smooth nuclear membrane without folds, nuclear pore, nucleus space uniform, evenly distributed nuclear chromatin, euchromatin-rich nucleus, occasionally nucleolus, cell quality rich in various organelles, normal morphology, structural integrity.Sham operation group:number of neurons and more occasional interstitial cell gap. The nuclei were round or oval, clear membrane structure, rules, uniform perinuclear space, nuclear chromatin distribution, occasionally heterochromatin. Abundant organelles, structural integrity, takes the shape of flattened Golgi vesicles can be seen parallel. Mitochondria were round, oval, film rules, uniform matrix; ribosome-rich endoplasmic reticulum. Model group:Number of neurons and more nuclei with irregular shape, decreased organelles, cell shrinkage, chromatin condensing different integrated block. Mitochondria decreased, swelling into a spherical, transparent into the vacuole; rough endoplasmic reticulum dilatation, off the surface of the ribosome. Apoptotic state of cells in more condensed cytoplasm, the whole cell electron density is higher, more irregular nuclei, chromatin highly condensed, marginalized.Acupuncture group:neuronal nuclear membrane more clear and complete, uniform chromatin, the cytoplasm shows a large number of mitochondria, were essentially normal, occasional swelling, cavitation; occasionally Golgi; endoplasmic reticulum rich in ribosomes, the cell has micro-swelling. Moxibustion group:a little fuzzy neuron membrane, chromatin uniform cytoplasm rich in organelles model group dropped out, showing that a large number of mitochondria, were essentially normal, small amount of swelling and cavitation. Endoplasmic reticulum rich in ribosomes, the cell has micro-swelling. Acupuncture+Moxibustion Group:neuronal membrane clear, complete and uniform chromatin, cytoplasm rich in organelles model group dropped out, showing that a large number of mitochondria were round, oval, film rules, more uniform matrix, occasional swelling; endoplasmic reticulum rich in ribosomes, the cell with micro-swelling.4. Monoamine neurotransmitters NE, DA,5-HT in brain tissues Normal control group and sham operation group rats brain NE, DA and 5-HT levels, no significant difference between (P> 0.05). Normal control group and model group rats'brain NE, DA and 5-HT content significantly between (P<0.05). And the normal control group has a higher content than the model group has. Model group and sham operation group rats'brain NE, DA and 5-HT content significantly between (P＜0.05). And sham operation group get a higher content than in the model group. Model group compared with the acupuncture group, moxibustion group, acupuncture+moxibustion group of rats'brain NE, DA and 5-HT levels were significant differences between (P<0.05). And acupuncture group, moxibustion group, acupuncture+moxibustion group has a higher content than the model group. Acupuncture group and acupuncture+moxibustion group of rats brain NE, DA and 5-HT content was no significant difference between (P>0.05).acupuncture combined with moxibustion moxibustion group and the group of rats brain Tissue NE, DA and 5-HT content was no significant difference between (P>0.05).5. Excitatory amino acid (Glu) and inhibitory amino acid (GABA) in the brain tissues Normal control group and sham operation group rats'brain, its Glu, GABA content, no significant difference between (P> 0.05). Normal control group and model group rats brain, its content of the difference was between significance (P<0.05). And the normal control group get a lower content of Glu but a higher content of GABA than the model group. Model group and sham operation group rats brain Glu content was significant difference between (P＜0.05), and the model group has a higher content than the sham operation group. Model group and sham operation group brain GABA content was significantly different between (P<0.05), and the model group was lower than the sham operation group.Conclusion:1. Acupuncture and moxibustion have a growth promoting effect on the CP rats which muscle strength and balance formed. There is almost no difference between them, thus get the nearly speed and magnitude of weight gain compared with normal and sham-operated rats. Simple acupuncture in this experiment was not found useful that further research is needed. A growth effect of strength and balance of CP rats are showed after having only moxibustion or acupuncture, but the better effect of acupuncture combined with moxibustion. Similarly, there is a great improvement of CP rats about the ability and intelligence after moxibustion and also have a better improvement of acupuncture combined moxibustion,2. In this study, compared with the model group, there is a larger amount of normal nerve cells in both moxibustion group, acupuncture group, acupuncture +moxibustion group and no significant difference between the three. It shows that acupuncture can inhibit to some extent in brain after cerebral hypoxia-ischemia, neuronal apoptosis, and hypoxic-ischemic brain injury has a protective effect, and the sooner the intervention the better.3. This study shows that a apoptosis of the neuronal cell in CP rats during transmission electron microscopy. It provides an experimental basis to the acupuncture+ moxibustion treatment of children with cerebral palsy that acupuncture treatment can reduce neuronal apoptosis, and thus to protect the nerve cells.4. Results:After needle or (and) after moxibustion treatment, content of DA, 5-HT, NE in brain of CP rats were significantly higher. Therefore, we can infer that:An increase content of monoamine neurotransmitters in brain tissue in rats treated with acupuncture may be the possible mechanism of brain paralysis5. In this study, Glu in brain tissue of rats in model group is significantly higher than those in sham operation operation group. Of which GABA is significantly lower than the sham operation group. Thus it is concluded that excitotoxicity of Glu and anti-excitotoxicity of GABA may be involved in pathophysiology of paralysis in the brain. Glu in brain tissues decreased significantly of treatment group when comparing with model group. It is showed that it can protect brain function by reducing Glu in ischemic brain of who got cerebral palsy by treating acupuncture, moxibustion or acupuncture combined with moxibustion therapy. GABA significantly increased in rats brain in Treatment group, indicating the acupuncture, moxibustion or acupuncture+moxibustion can increase the content of GABA against Glu excitotoxicity induced, and the effect among the non-significant difference. Therefore, the process in the treatment of CP, the maintenance of excitatory and inhibitory neurotransmitter neurotransmitter homeostasis may play a role in the mechanism of acupuncture.
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