| BackgroundNeonatal hypoxic-ischemic brain damage (HIBD) is due to asphyxia caused cerebral blood supply and gas exchange impairment caused by a brain injury, a serious disease endangering the health of newborn lives, have a higher morbidity and mortality, also one of the important reasons contributed to children's mental retardation, cerebral palsy and other neurological disabilities. The causes and pathogenesis of HIBD are more complex; more and more evidence has showed that apoptosis is also involved and plays a unique role. The occurrence of neuronal apoptosis and prognosis in children is closely linked. At the molecular level, apoptosis is affected by many genes and the regulation of cytokines, nuclear factor-κB and caspase-3 are the research focus.Acupuncture and moxibustion are the important part of traditional Chinese medicine.They are very useful for children with cerebral palsy. We have did a lot of work in clinical research, but a little work in experiment research. We could not understand clearly the specific mechanism of acupuncture and moxibustion. For of all the reasons, our team investigated the effect of acupuncture and moxibustion under governor vessel treatment on movement function, balance function, study and memery function, and brain NF-Kb, Caspase-3 expressing in the HIBD rats. Attempts to explore the mechanism of acupuncture and moxibustion for hypoxic-ischemic brain damage from various aspects.ObjectiveTo investigate the effect of acupuncture and moxibustion under governor vessel treatment on movement function, balance function, study and memery function, and brain NF-Kb,Caspase-3 expressing in the CP rats. Meanwhile, to understand the mechanism of early acupuncture and moxibustion treatment for CP, and to provide the theoretical basis for the promotion of early acupuncture and moxibustion treatment in clinical use for children with cerebral palsy.Methods1. Established a simulation model of perinatal asphyxia HIBD2. The animals were randomly divided into five groups:normal group, sham group, model group, simple acupuncture group and acupuncture with moxibustion group. We developed a treatment timetable with acupoints reference from (Study of experimental acupuncture), combining human and animal bones analogy: acupoints taken for treatment include parietal bone center for'Baihui' external ear canal, straight 0.8cm simulate take'Temporal 1 needle'; radial bone proximal lateral aspect from the wrist off 3cm or so take'Neiguan' radius proximal lateral joint when elbow is flexed take'Quchi'; anterior aspect of lower leg under the fibular head and about 5mm take'Zusanli' hind legs before the middle of the sole take'Yongquan'. After treatment for two stages, separately carry out tests for HIBD mouse model on behavior motor ability and on learning and memory ability, and then after mice were sacrificed, examine the model mouse brain tissue and related gene expression targets.3. Hanging test, Slope test and Y maze test were used for valuing neuroethology function in different groups.The method of immunohistochemistry was used to observe the HIDB mouse dynamic expression of apoptosis regulatory proteins NF-Kb and Caspase-3.Results1. Modeling resultsMortality rate:33.3%, Success rate:61.11%2. Change of weightMultiple comparison results showed that:the normal group, sham operation group, acupuncture group, acupuncture and 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.Before intervention, there were no significant difference among five groups. After intervention, the weight of rats were heavier than before. Model group was lighter than other groups. From heavy to light, the order was normal group, sham operation group, acupuncture and moxibustion group, acupuncture group and model group.3. Hanging testBefore intervention, model group, acupuncture group and acupuncture with moxibustion group were worse than normal group and sham operation group.After the first treatment, the situation was the same as before., but acupuncture with moxibustion group was better than acupuncture groupAfter the second treatment, model group, acupuncture group and acupuncture with moxibustion group were also worse than normal group and sham operation group. From better to worse,the order was acupuncture and moxibustion group, acupuncture group and model group.After intervention, the valu of Hanging test were better than before in five groups.4. Slope testBefore intervention, model group, acupuncture group and acupuncture with moxibustion group were worse than normal group and sham operation group.After the first treatment, the situation was the same as before., but acupuncture with moxibustion group was better than acupuncture groupAfter the second treatment, model group, acupuncture group and acupuncture with moxibustion group were also worse than normal group and sham operation group. From better to worse, the order was acupuncture and moxibustion group, acupuncture group and model group.After intervention, the valu of Slope test were better than before in five groups.5. Y maze testThe times of right movement, there were significant difference among five groups. From better to worse, the order was acupuncture and moxibustion group, acupuncture group, model group, normal group and sham operation group.6. Pathological changes in brain tissueHE staining:normal light microscope under 10X20 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, hippocampal 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 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.7. Counting of normal nerve cells in brain cortexModel group was worse than normal group and sham operation group.Acupuncture with moxibustion group and acupuncture group were better than model group. There were no significant difference between acupuncture with moxibustion group and acupuncture group.8. Expressing of proteins NF-ΚbModel group was worse than normal group and sham operation group.Acupuncture with moxibustion group and acupuncture group were better than model group. There were no significant difference between acupuncture with moxibustion group and acupuncture group.9. Expressing of proteins Caspase-3Model group was worse than normal group and sham operation group.Acupuncture with moxibustion group and acupuncture group were better than model group. There were no significant difference between acupuncture with moxibustion group and acupuncture group.Conclusion1. Hypoxic-ischemic brain injury in rats caused neural cell apoptosis. There were clsed relationship between neural cell apoptosis and expressing of proteins NF-Κb and Caspase-3.2. The therapy of early combining acupuncture with moxibustion was useful for the improvement of neural function, and acupuncture with moxibustion group was better than acupuncture group.3. The therapy of early combining acupuncture could improve the pathological changes in brain tissue.4. The therapy of early combining acupuncture has positive regulation in the expressing of proteins NF-Kb and Caspase-3. |
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