| Objective:Intracerebral hemorrhage is a serious threat to human health and high mortality. Functional deterioration of the nerves following cerebral hemorrhage is associated with cerebral hemorrhage induced brain edema and involves mutiple pathways of nerve injuries. Such as space-occupying effect, blood flow changes in tissues surrounding the hematoma and secondary ischemia, secondary edema following cerebral hemorrhage, clot retraction and hydrostatic pressure, coagulation cascade and thrombin production, erythrocyte lysis and hemoglobin toxicity, rupture of the blood-brain barrier, activation of complement, inflammation and neuronal apoptosis after cerebral hemorrhage.Brain edema and inflammatory response have become hot topic. Aquaporin-4(AQP-4) is mainly distributed in brain tissue.It is a key channel for water flow in the brain and a indicator of intracerebral edema. Tumor Necrosis Factor(TNF-α) and Interleukin-10(IL-10) are important inflammatory cytokines.They can react inflammatory response in some ways. Nerve growth factor(NGF) plays an extremely important role in the development and survival of nerve cells.It prevent atrophy and die of the neurons during brain injury and promote restoration of the neurons. Its role is mainly reflected in the maintenance of neuronal survival; promoting axonal regeneration and determining the direction of their growth; making axons located in the appropriate target cells to form functional connections; promoting myelination.The purpose of this experiment is to explore the changes of AQP-4, TNF-α and IL-10 after injecting NGF into the hematoma cavity of intracerebral hemorrhage in rats. Providing the basis for intracranial hematoma aspiration combine with NGF in clinical.Methods: 144 healthy male SD rats which weight are between 290 g and 310 g, are randomly divided into four groups:ICH model group,saline treatment group, dexamethasone treatment group and NGF treatment group. There are 36 rats in each group. Among these 18 rats are for the determination of brain water content. Fixing the rats on the stereotactic instrument after the rat were anaesthetized. Injecting non-heparinized autologous arterial blood into the right caudate nucleus to making for rat model of intracerebral hemorrhage. Each group of rats are injected 60μl autologous arterial blood. Selecting qualified rats according to the Longa standard after 24 hours. The intracerebral hemorrhage model group is not given drug treatment.The saline group are injected saline at the same site in the hematoma cavity according to 0.2ml/kg. The dexamethasone treatment group are injected dexamethasone according to 1ml/kg. The NGF treatment group are injected NGF according to 1.67 ×10-3AU/ kg. Choose six rats at 48 hours, 3 days and 7 days respectively. Take hematoma surrounding brain tissue and then cut into 6 slices to observe AQP-4, IL-10 and TNF-α expression by immunohistochemical methods. Take six high-power field in each section and calculate the positive express. Cut off the head of three rats and get the brain at three time points, then measure wet weight and dry weight. Calculate the water content of the brain tissue by dry-wet method. And then use SPSS 13.0 statistical data to observe the alteration of each indicator at different phase from the hematoma surrounding brain tissue.Result:1 The Changes of the Water Content of the Rat Brain TissueThe brain water content of the ICH model group,the saline group, the dexamethasone group and the NGF group increase at 48 hour, reach the peak on the third day after ICH and decrease on the seventh day. The ICH model group and the saline group have no statistically significant differences at the 3 day(P>0.05). The NGF group and the dexamethasone group have no statistically significant differences at the 48 hour(P>0.05). There are significant differences among the remaining groups(P<0.05).2 The Changes of the AQP-4 ExpressionThe dexamethasone group compare with the NGF group. The two groups have no statistically significant differences at the 48 hour and 7 day(P>0.05).The ICH model group and saline group compared with the dexamethasone group and NGF group.the expression of AQP-4 increase at 48 hour, 3 day and 7 day after ICH have a statistically significant difference(P<0.05). It increase at 48 hour,reach the peak on the third day and decline on the seventh day.3 The Changes of the TNF-α ExpressionThe NGF group, the dexamethasone group, the saline group and ICH model group all increase at 48 hour, 3 day and 7 day after ICH have a statistically significant difference(P<0.05). The expression of TNF-α begin to increase in 48 hours, peak on the third day and decline on the seventh day after ICH.4 The Changes of the IL-10 ExpressionThe dexamethasone group compare with the NGF group.The two groups have no statistically significant differences at the 48 hour, 3day and 7 day(P>0.05).The ICH model group and saline group compared with the dexamethasone group and NGF group. The expression of IL-10 increase at 48 hour, 3 day and 7 day after ICH have a statistically significant difference(P<0.05). The expression of IL-10 begin to increase at 48 hour peak on the third day and decline on the seventh day after ICH.Conclusions:1 The method of rat autologous arterial blood injection produce intracerebral hemorrhage model have higher successful rate.2 The expression of AQP-4 and water content around the hematoma have a positive relationship.3 The expression of TNF-α after ICH involve in brain tissue necrosis, and is related to cerebral edema.4 IL-10 mediated anti-inflammatory response, increased rapidly in a short time, lasted for a long time, less volatile.5 NGF is injected into the hematoma cavity can reduce the content ofintracerebral edema. Perihematomal AQP-4, TNF-α expression decrease. It regulate IL-10 expression around hematoma. It show that NGF can reduce edema, inhibit the release of pro-inflammatory cytokines and regulate the expression of anti-inflammatory cytokines. Provide an experimental basis for clinical. |
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