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目的观察褪黑素(Mel)对癫大鼠海马神经元凋亡及半胱氨酸蛋白酶(caspase)-3表达的影响。方法采用匹罗卡品(Pilo)制作大鼠癫持续状态(SE)模型,随机分为Pilo组、Mel组和对照组,用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)染色和免疫组化技术检测大鼠海马神经元凋亡数和caspase-3的表达,并与对照组比较。结果SE后6h,Pilo组开始出现少量TUNEL阳性细胞;SE后72h,达到高峰;SE后7d,TUNEL阳性细胞开始减少。SE后6h,Pilo组大鼠海马caspase-3阳性细胞数增多,主要集中于CA1和CA3区;SE后48h,达到高峰;SE后72h,阳性细胞数开始减少;SE后7d,caspase-3表达基本恢复正常。Mel组各时间点大鼠海马TUNEL阳性细胞数和caspase-3表达均明显低于Pilo组大鼠(均P<0.01)。结论Mel可减少癫大鼠海马神经元凋亡,抑制caspase-3的表达,起到神经保护作用。
Objective To investigate the effect of melatonin (Mel) on the neuronal apoptosis and the expression of caspase-3 in the hippocampus of epileptic rats. Methods The model of epileptic seizure (SE) in rats was established by Pilo. The model rats were randomly divided into Pilo group, Mel group and control group. The terminal deoxynucleotidyl transferase mediated dUTP nick end labeling TUNEL staining and immunohistochemistry were used to detect the number of apoptotic hippocampal neurons and the expression of caspase-3 in hippocampus of rats and compared with the control group. Results Six hours after SE, a small amount of TUNEL-positive cells began to appear in Pilo group; the peak appeared at 72 hours after SE, and TUNEL-positive cells began to decrease at 7 days after SE. At 6h after SE, the number of caspase-3 positive cells in hippocampus of Pilo rats increased, mainly in CA1 and CA3 areas; the peak appeared at 48h after SE; the number of positive cells began to decrease at 72h after SE; the expression of caspase-3 The basic return to normal. The number of TUNEL positive cells and the expression of caspase-3 in hippocampus of Mel group were significantly lower than those of Pilo group at each time point (all P <0.01). Conclusion Mel can reduce the apoptosis of hippocampal neurons in epileptic rats and inhibit the expression of caspase-3, which plays a neuroprotective role.