| Neonatal hypoxic-ischemic brain damage (hypoxia-ischemic brain damage, HIBD) is the most common causes to neonatal deaths and the neonatal nervous system damage.According to statistics, there are about 1,2 HIBD neonatal cases caused by perinatal asphyxia in 1000 cases.There are about 15～20% children with HIBD will dead in the neonatal period, approximately 25% of the surviving children are left with neurological damage, such as cerebral palsy, mental retardation, epilepsy and so on.Neonates with HIBD are mainly with symptomatic and supportive treatment hypothermia and hyperbaric oxygen therapies have been widely recognized, The effect of hypothermia therapy on moderate HIBD significant, and can significantly reduce the occurrence and prevention of complications in children, improve their quality of lives.20 years ago,there is generally accepted that the brain tissue after the fully developed nerve cells almost no ability to regenerate not like as the regeneration of other rgans.But recent studies found that there are neural stem cells (NSC) in some special areas,and the brain brain tissue has the neural regeneration ability.Neural stem cells are some cells which have the ability of self-renewal and multilineage differentiation potential in embryonic and adult neural tissues. They can differentiate into neurons, astrocytes and oligodendrocytes, and participate in neural development and nerve injury repair. In addition, recent studies have shown that there are many inhibitors of central nervous system, peripheral nerve axons of adult mammals can be widely regeneration after injury, but the capacity of CNS axon regeneration is limite, its reasons not only because damage points in addition to the lack of growth-promoting molecules, damage intrinsic growth capacity of neurons and glial scar poor physical barrier,but also the micro-environment in the presence of inhibitory factor plays an important role, such as Nogo-A.It is an important hub of axonal growth inhibitory factor, its central nervous system Inhibition of injury has been confirmed in vivo experiments. The early brain injury of children with relatively large plasticity, so researchs to maximize the stimulation of endogenous neural stem cells produce and promote their proliferation and differentiation are a hotspot in recent years.ObjectiveThis experiment produce the neonatal hypoxic-ischemic brain injury rat model, and the treatment the neonatal hypoxic-ischemic brain injury rat model with the Scalp Sites Medication Injection.So to analysis the effect of Scalp Sites Medication Injection on hypoxic-ischemic brain damage (HIBD) neonatal rats of endogenous neural stem cells Proliferation, differentiation, and inhibition of nerve regeneration factor Nogo-A mRNA expression.MethodsSeven-day-old SD rats were randomly divided into 4 groups:sham operation group, model of HIBD group, sites injected with normal 9g/L group, and sites injected with VitB1,VitB12 group.Rat model of HIBD was prepared by ligation of left common carotid artery and later with a temporary systemic hypoxia (80mL/L O2) for 2 hours. Fourteen days later, sites injected with physiological saline group and the sites injected with VitB1,VitB12 group were respectively treated with physiological saline and mixed diluent of VitB1,VitBi2 in rats left frontal lobe and apieal lobe 1 time one day, for 20 days. Sham operation group and model group were not treated. After the intervention, about 7 days later, all groups of rats were killed:brains were removed after cardiac perfusion organizations with 40g/L paraformaldehyde, and separately continuous slices.Using the HE stain for pathological analysis.The expression of Nestin using as neural stem cell marker in hippocampus was determined by immunofluorescence,and detected the expression of glial fibrillary acidic protein (GFAP) and neuron specific enolase (NSE) with immunohistochemisty. Determinated the expression of nerve regeneration inhibitory factor Nogo-A mRNA by in situ hybridization.Using SPSS 16.0 statistical package to analyze, measurement data were demonstrated by mean±standard deviation, using one-way ANOVA to compare the mean of each group,α=0.05 is the significance level.Results1.HE staining showed that:site of injection can reduce neuronal degeneration and necrosis and promote their rehabilitation and repair,and the effect is more apparent on the sites injected with group.2.Sites medication injection can increase the expression of Nestin in hippocampus, which with the highest expression by injected with VitB1,B12 but with the lowest expression for the sham operation group.3.NSE Immunohistochemistry showed that the sham operation group and sites injected with VitB1,B12 group expressed higher than others, and the difference between them was small, but the model group had the least expression.(P<0.005).4.The expression of GFAP immunohistochemistry in hippocampus showed that the sham operation and the model group were respectively with the least and the most expression.Sites injected withphysiological saline and sites injected with VitB1 VitB12 can inhibit its expression,which is obviously for sites injected with VitB1,VitB12 group (P<0.005).5.The expression for sham operation group of Nogo-A mRNA was at the least,but the expression of model group was at the opposite way.Sites injected with with physiological saline and sites injected with VitB1,VitB12 groups had a lower expression than the model group,which was obviously for sites injected with VitB1,VitB12 group (P<0.005).ConclusionsScalp sites medication injection proved the performance of rats with cerebral palsy, and reduce nerve injury.The group using scalp sites medication injection increased the expression of Nestin and NSE, but GFAP and Nogo-A mRNA expression is reduced. This suggests that scalp sites injection can promote endogenous neural stem cell proliferation and differentiation, inhibition of Nogo-A mRNA expression, and promote nerve regeneration and repair,and joint use of medication, can enhance the treatment effect. |
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