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目的探讨巨细胞病毒对乳鼠海马神经元N-甲基-D-天冬氨酸(NMDA)受体NR1亚基mRNA表达水平的影响。方法建立乳鼠海马神经元巨细胞病毒感染细胞模型,以感染后8、16、24h三个不同的时间点随机分为病毒组和对照组,观察海马神经元形态学改变,同时采用实时定量PCR技术检测病毒对海马神经元NMDA受体NR1亚基mRNA表达水平的影响。结果与对照组比较,病毒组逐渐出现细胞病变效应,最后细胞死亡;病毒组循环域值(Ct)8h为35.75±0.096、16h为34.636±1.565、24h为36.95±1.638。而对照组Ct值分别是30.60±0.37、28.937±0.064、36.95±1.638。两组差异有统计学意义(均P<0.05),病毒组内各时相目的基因两两比较F=130.629,P=0.00001。差异有统计学意义。提示病毒组NMDA受体NR1亚基mRNA表达水平均下降,并呈一定的时间-效应关系。结论巨细胞病毒对乳鼠海马神经元形态学发育和NMDA受体NR1亚基mRNA表达水平有严重影响,这可能是小鼠巨细胞病毒宫内感染导致学习记忆能力发育落后的分子机制之一。
Objective To investigate the effects of cytomegalovirus on the expression of N-methyl-D-aspartate (NMDA) receptor NR1 subunit mRNA in neonatal rat hippocampal neurons. Methods To establish a cell model of cytomegalovirus infection in hippocampal neurons of suckling mice. The infected cells were randomly divided into virus group and control group at different time points of 8, 16 and 24 hours after infection. The morphological changes of hippocampal neurons were observed. Real-time quantitative PCR Effect of Virus Detection on NR1 Subunit mRNA Expression in Hippocampal Neurons. Results Compared with the control group, the cytopathic effect was gradually observed in the virus group, and finally the cell death was observed. The Ct value of the virus group was 35.75 ± 0.096 at 8h, 34.636 ± 1.565 at 16h, and 36.95 ± 1.638 at 24h. While the control group Ct values were 30.60 ± 0.37,28.937 ± 0.064,36.95 ± 1.638. The difference between the two groups was statistically significant (P <0.05). The comparison of the target gene in each group in each group was F = 130.629 (P = 0.00001). The difference was statistically significant. Suggesting that NR1 subunit NMDA receptor mRNA expression in the virus group were decreased, and showed a time-dependent relationship. Conclusion Cytomegalovirus has a serious impact on the morphological development of hippocampal neurons and the expression of NR1 subunit mRNA of neonatal rat hippocampus, which may be one of the molecular mechanisms of intrauterine cytomegalovirus infection that leads to poor development of learning and memory.