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目的观察神经生长因子(NGF)对红藻氨酸(KA)致大鼠海马半胱氨酰天冬氨酸蛋白酶-3(Caspase-3)表达的影响,探讨其对癫大鼠神经细胞的保护作用及其机制。方法将60只日龄21～30 d健康大鼠随机分为9 g.L-1盐水对照组(A组),KA致模型对照组(B组),NGF治疗组(C组),每组20只。KA用9 g.L-1盐水配成2 g.L-1,B组、C组大鼠以4 mg.kg-1腹腔注射。注射完观察记录其性发作的潜伏期和性发作的级别,并进行EEG和病理学检查。C组每只大鼠腹腔注射KA20～30 min予NGF4μg.kg-1腹腔注射。A组用9 g.L-1盐水2 mL.kg-1灌胃和腹腔注射。分别于KA注射6 h、1 d、3 d时将各组大鼠处死5只,常规方法灌注固定,制作冷冻切片。用免疫组织化学染色SP法检测大鼠海马Caspase-3表达,并用HPIAS-2000显微图像定量分析系统进行定量分析。结果B组、C组大鼠注射KA后30 min左右开始出现性发作,EEG和病理学检测均显示癫发作特征。C组大鼠注射3 d后,Caspase-3阳性神经元计数为7.10±2.80,B组为18.40±3.86;A组为0.34±0.06。C组大鼠海马Caspase-3阳性神经元计数与B组相同时间点比较均减少,差异有统计学意义(Pa<0.05);C组大鼠海马Caspase-3阳性神经细胞平均灰度值各时间点均小于B组;C组注射3 d后,灰度值为90.64±4.24,B组为210.95±6.25,二组比较差异有统计学意义(P<0.05)。结论NGF能抑制癫大鼠海马Caspase-3表达,对癫发作大鼠具有神经细胞保护作用。
Objective To investigate the effect of nerve growth factor (NGF) on the expression of caspase-3 in hippocampus of rats induced by kainic acid (KA) Protection and its mechanism. Methods Sixty healthy, aged 21-30 days old rats were randomly divided into 9 g L-1 saline control group (group A), KA model group (group B) and NGF treatment group (group C) only. KA with 9 g.L-1 saline dubbed 2 g.L-1, B group, C group 4 mg.kg-1 intraperitoneal injection. After injection, observe the latency of episodes and the grade of seizures, and perform EEG and pathological examination. Rats in group C were intraperitoneally injected with KA for 20-30 min to give intraperitoneal injections of NGF 4 μg.kg-1. A group with 9 g.L-1 saline 2 mL.kg-1 intragastric and intraperitoneal injection. Five rats were sacrificed at 6 h, 1 d and 3 d after KA injection respectively. The rats were perfused and fixed by conventional method to make frozen sections. The expression of Caspase-3 in rat hippocampus was detected by immunohistochemical SP method and quantified by HPIAS-2000 micrograph quantitative analysis system. Results In group B and group C, the onset of seizures began 30 minutes after KA injection, and both EEG and pathological examination showed the characteristics of epileptic seizures. After 3 days of injection, the number of Caspase-3 positive neurons in group C was 7.10 ± 2.80, in group B was 18.40 ± 3.86 and in group A was 0.34 ± 0.06. The number of Caspase-3 positive neurons in hippocampus of C group decreased compared with that of B group at the same time point, the difference was statistically significant (Pa <0.05); the mean gray value of Caspase-3 positive neurons in hippocampus of C group was (P <0.05). After 3 days of injection, the gray value of group C was 90.64 ± 4.24 and that of group B was 210.95 ± 6.25. The difference between the two groups was statistically significant (P <0.05). Conclusion NGF can inhibit the expression of Caspase-3 in the hippocampus of epileptic rats and protect the rats from epileptic seizures.