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目的观察孕早期大鼠人巨细胞病毒(HCMV)感染对仔鼠海马细胞外信号调节蛋白激酶(ERK)通路的影响,为研究HCMV先天性感染对子代大鼠海马学习记忆损伤的机制及防治HCMV先天性感染导致的学习记忆障碍等神经系统后遗症提供实验依据。方法取8～9周龄雌性SPF级SD大鼠12只,按雌雄=31配对,孕鼠随机分为对照组和实验组,每组6只。于孕第5天实验组孕鼠腹腔内注射100半数组织细胞感染量HCMV悬液0.8 mL,对照组孕鼠腹腔内注射等量的人胚肺成纤维细胞上清液。于出生后30 d每组随机取10只仔鼠,取仔鼠脑组织,采用HE染色观察仔鼠海马齿状回(DG)区颗粒细胞的形态学改变;免疫组织化学法观察仔鼠海马DG区ERK1、ERK2、p-ERK的表达,并应用SPSS 13.0软件进行分析。结果对照组仔鼠海马未见明显的神经元缺失,排列紧密,均匀整齐,胞核呈圆形或椭圆形,核仁明显,染色质均匀;实验组仔鼠海马细胞数减少,组织结构较稀疏,排列紊乱。ERK1、ERK2、p-ERK主要表达于胞核和胞质,阳性部位呈现棕黄色。实验组仔鼠海马DG区的ERK1、ERK2、p-ERK染色强度及平均积分光密度均低于对照组,差异均有统计学意义(Pa<0.01)。结论孕早期HCMV感染可降低仔鼠海马DG区ERK1、ERK2、p-ERK的表达,可能与HCMV先天感染损伤海马的学习记忆功能有关。
Objective To observe the effect of HCMV infection in early pregnancy on the extracellular signal-regulated kinase (ERK) pathway in hippocampus of neonatal rats. To study the mechanism and prevention and cure of congenital HCMV infection in hippocampal learning and memory of offspring HCMV congenital infection caused by learning and memory disorders and other nervous system sequelae provide experimental evidence. Methods Twelve male Sprague-Dawley (SD) rats aged 8 to 9 weeks were paired with female mice at the ratio of 3 to 1. The pregnant rats were randomly divided into control group and experimental group, with 6 rats in each group. On the fifth day of pregnancy, pregnant mice in the experimental group were injected intraperitoneally with 0.8 ml HCMV suspension of 100 cells, and the control group was intraperitoneally injected with the same amount of human embryonic lung fibroblast supernatant. At 30 days after birth, 10 offspring were randomly selected from each group, and the brain tissues of the offspring were taken. The morphological changes of granulosa cells in the hippocampal dentate gyrus (DG) were observed by HE staining. The hippocampal DG The expression of ERK1, ERK2 and p-ERK was analyzed by SPSS 13.0 software. Results The hippocampus of the control group showed no obvious neuron loss, arranged closely, uniformly and neatly, the nucleus was round or oval, the nucleoli were obvious and the chromatin was even. The number of hippocampus in the experimental group was decreased and the structure was sparse , Arranged disorder. ERK1, ERK2, p-ERK mainly expressed in the nucleus and cytoplasm, the positive part showed brown yellow. The staining intensity and average integrated optical density of ERK1, ERK2 and p-ERK in hippocampus DG of experimental group were lower than that of control group (P <0.01). Conclusion HCMV infection in early pregnancy can reduce the expression of ERK1, ERK2 and p-ERK in hippocampus DG of neonatal rats, which may be related to learning and memory function of HCMV congenital infection in hippocampus.