The Effects Of Intracerebroventricular Injection Of P63 Antibody On P63 And P53 Expression And Neuronal Apoptosis After Cerebral Ischemia In Rats

Objective:To observe the effects of intracerebroventricular injection of P63 antibody on P63 and P53 expression, neuronal apoptosis in hippocampal CA1 area after global cerebral ischemia in rats, and to explore the effects of p63 on P63 and P53 expression and neuronal apoptosis after cerebral ischemia and anoxia, and to provide new theoretical basis for prevention and cure of neuronal apoptosis after cerebral ischemia and anoxia.Methods:According to random number table,75 male SD rats were randomly divided equally into three groups:group A, B and C. The rats in group A were received artificial cerebrospinal fluid injected into lateral ventricle without cerebral ischemia; the rats in group B were received artificial cerebrospinal fluid injected into lateral ventricle and cerebral ischemia; the rats in group C were received P63 antibody injected into lateral ventricle and cerebral ischemia. The neurological behavior scores of rats in three groups were evaluated before anesthesia, and the rat with 16-20 scores was selected into this study. All rats were evaluated espectively neuroethology at 6h,12h,24h,48h,72h after operation. Under general anesthesia, all rats were executed and brain tissues were collected after perfusion with 4% formaldehyde polymerisatum. The neuronal densities were determined by HE, neuronal apoptosis indexes were determined by TUNEL, P63 and P53 expression were detected by immunohistochemistry (IHC) and semi-quantitative analysis in hippocampal CA1 area.Results:1.Animal model In the study,7 rats died:5 rats died from cardiac arrest and apnea, and 2 rats died after operation. Seven rats were complemented. Seventy-five rats were enrolled in the statistical analysis.2.Neurological behavior scores Neurological behavior scores of 75 rats were normal before operation(16^20 points). The scores were normal in group A at each time point after operation. In group B and C, postoperative neurological behavior scores were significantly lower than preoperation (p<0.01), and the scores were lowest at 72h after operation (p<0.05). But postoperative scores were no significantly difference between group B and C at 6h after operation (p> 0.05), and the scores in group C were significantly higher than group B at other time points(p<0.01).3.Neuronal density The neuronal densities in hippocampal CA1 area were no significantly difference at each time point after surgery in group A (p>0.05). The neuronal densities in group B and C were significantly lower than group A at each time point after operation (p<0.021) and lowest at 72h (p<0.05). But the neuronal densities in group C were significantly higher than group B at all time points after operation (p<0.05).4.Neuronal apoptosis Apoptotic neurons were very rare in hippocampal CA1 area of group A at all time points after operation. In group B and C, neuronal apoptosis indexes in hippocampal CA1 area were significantly higher than group A at each time point after operation(p<0.01), and the indexes were highest at 48h(p<0.05). But neuronal apoptosis indexes in group C were significantly lower than group B (p<0.05).5.P63 and P53 expression P63 and P53 expression in hippocampal CA1 area of group A were rare at all time points after surgery. P63 and P53 expression in hippocampal CA1 area in group B and C were significantly higher than group A at all time points after operation (p<0.01). P63 expression began to increase at 6h after surgery, highest at 48h (p<0.05) after surgery and then its expression reduced. And P53 expression started to rise at 6h after operation, highest at 24h and then reduced gradually. There were no significant differences in P53 expression between group B and C at each time point after surgery(p>0.05), but P63 expression in group C were significantly lower than group B(p<0.05).6.Correlation analysis The expression of P63 had positive correlation with P53 expression in hippocampal CA1 area after cerebral ischemia in rats (r=0.592,p<0.01); and there was a positive correlation between P63 expression and neuronal apoptosis in hippocampal CA1 area after cerebral ischemia (r=0.853, p<0.01); and P53 expression was positive correlated with neuronal apoptosis after cerebral ischemia (r=0.852,p<0.01).Conclusion:1. Both P63 and P53 expression increased significantly in hippocampal CA1 area after cerebral ischemia in rats, and intracerebroventricular injection of P63 antibody could reduce P63 expression, but had no significantly effect on P53 expression.2. Intracerebroventricular injection of P63 antibody could reduce neuronal apoptosis after cerebral ischemia in rats, and improve neurological behavior scores, and therefore could have neuroprotective effect.3. There were positive correlations among P63, P53 and neuronal apoptosis in hippocampal CA1 area after cerebral ischemia in rats.