| Background and ObjectiveIntracerebral hemorrhage (ICH) is an emergent disease. At present, there are many research results reveal that cell apoptosis participates in the secondary injury of neurocytes. So cell apoptosis is an important factor of neurocytes injury of intracerebral hemorrhage. Compared with non-diabetics, diabetics have a worse hospital and long-term outcome after intracerebral hemorrhage. Hyperglycemia may promote the apoptotosis of neurons, which has been verified in the ischemic cerebrovascular disease. At present, it was researched rarely in the ICH.In the study, using immunohistochemistry and TUNEL methods, the neuron apoptosis and the expression of Bax and Bcl-2 were explored in rat ICH model and hyperglycemia model, which may provide strategies for clinical treatment on cerebral hemorrhage.Materials and methodsNinety-six healthy adult male SD rats were randomly divided into 4 groups: sham group, normal plasma glucose group, hyperglycemia group and insulin intervention group. Each group was divided into 4 various stages:6h,24h,3d,7d.Hyperglycemia model was induced by the injection of streptozotocin (STZ) into abdominal cavity by injection syringe. The model was successful until blood glucose was higher thanl1.1mmol/L.ICH model was induced by the injection of autologous unaggregated artery blood into the right basal ganglia of rats by stereotaxic apparatus.Preparation of the tissue section:The successful models were perfused and took tissue at the according time. The tissue was taken coronary sections before and behind the pin hole.Examine parameter:Bax and Bcl-2 were studied with immunohistochemistry. Positive area average integral optical density value was measured with image analysis system, its mean value was average integral photodensity. Nerve cell apoptosis was measured by terminal-deoxynucleotidyltransferase mediated nick end label (TUNEL) method. Apoptotic index (AI)=apoptotic neurons/total neurons×100 %.Statistic analysis:All data were expressed as mean±SD, and analyzed with the SPSS13.0 software, the means of the samples were compared using one-way analysis of variance followed by Least Significant Difference (LSD) for multiple comparisions. Correlation between Bax and TUNEL was judged by linear regression. The significant difference was judged by a=0.05.Results1. The dynamic changes of Bax and Bcl-2The expression of Bax and Bcl-2 in the striatum of sham group rats was low and changed a little from 6h to 7d. The expression of Bax and Bcl-2 in the perihematomal areas at different time points after ICH in ICH group, hyperglycemia group and insulin intervention group began at 6h and increased gradually, ultimate in 24h, still can be detected at 7d. The expression of Bax and Bcl-2 in the hyperglycemia group was significantly higher than that in the other groups at the same time point (P<0.05), the above-mentioned index of insulin intervention group were lower than that in hyperglycemia group and higher than that in the sham group at the same time point (P<0.05), and had no obvious difference with that in the normal glucose group at the same time point (P>0.05).2. The dynamic changes of neuron apoptosisApoptotic neurons were few in the striatum of of sham group rats, and AI had small change from 6h to 7d. AI in the perihematomal areas at different time points after ICH in ICH group, hyperglycemia group and insulin intervention group began at 6h and increased gradually, ultimate in 24h, still can be detected at 7d. AI in the hyperglycemia group was significantly higher than that in the other groups at the same time point (P<0.05), the above-mentioned index of insulin intervention group were lower than that in hyperglycemia group and higher than that in the sham group at the same time point (P<0.05), and had no obvious difference with that in the normal glucose group at the same time point (P>0.05).3. The result of the relationship analysis of apoptotic and expression of BaxAI positively correlates with the expression of Bax in hyperglycemia group at the same time point(r=0.80, P<0.05).Conclusion1. Hyperglycemia increases the apoptotic neurons and stimulates the expression of Bax and Bcl-2 in perifocal tissue of intracerebral hemorrhage in rats.2. Apoptotic index positively correlates with the expression of Bax in hyperglycemia group at the same time point.3. Control of blood glucose level with insulin can reduce neuron apoptosis.
|
PDF Full Text Request