Yuan Tian-Ming, Yu Hui-Min, Gu Wei-Zhong, Li Jian-Ping
Department of Neonatology, Laboratory, Children's Hospital, Zhejiang University School of Medicine, Hangzhou, PR China.
J Perinat Med. 2005;33(5):415-22. doi: 10.1515/JPM.2005.074.
In order to investigate the neuropathological effects on the developing rat brain after intrauterine infection, identification of glail fibrillary acidic protein (GFAP), 2', 3'-cyclic nucleotide phosphodiesterase (CNPase), and neurofilament (NF) was observed. Escherichia coli (E. coli) was inoculated into uterine horn of pregnant rats when gestation was 70% complete (15 days) and the control group was inoculated with normal saline. Immunohistochemistry was used for evaluation of GFAP, CNPase, and NF expression in pup brains at postnatal day 7 (P7) and reverse transcriptase-PCR (RT-PCR) to analyze macrophage inflammatory protein-1 alpha mRNA (MIP-1 alpha mRNA), macrophage inflammatory protein-1 beta mRNA (MIP-1beta mRNA), the regulated upon activation normal T expressed and secreted chemokine mRNA (RANTES mRNA) and Eotaxin mRNA expression in pup brains at P1, P3 and P7. The numbers of GFAP-positive cells of the E. coli-treated group pups were marked increased in periventricular white matter and hippocampus at P7 compared with the control group but no significant different levels of GFAP expression in corpus callosum were found between two groups. The integrate density (ID) of CNPase-positive staining of the Escherichia coli-treated group pups were marked decreased in periventricular white matter and corpus callosum at P7 compared with the control group. The ID of NF-positive staining of the Escherichia coli-treated group pups were marked decreased in periventricular white matter at P7 compared with the control group and no significant different levels of NF expression in corpus callosum were found between two groups. The expression of MIP-1 alpha mRNA and MIP-1 beta mRNA in brain of the E. coli-treated pup rat were higher than the control at P1, but the expression of MIP-1 alpha mRNA and MIP-1 beta mRNA in brain of the pup rat at P3 and P7 had no significant difference between two groups. The alteration of expression of GFAP, CNPase, and NF in the brain of neonatal rats after intrauterine infection suggested that intrauterine infection could cause neonatal white matter damage. Moreover, the transient increase in expression of chemokine such as MIP-1 alpha, MIP-1 beta in neonatal brain after intrauterine infection indicated that MIP-1 alpha, MIP-1 beta may be a mechanism mediating between the neonatal white matter damage and the intrauterine infection.
为了研究宫内感染后对发育中大鼠脑的神经病理学影响，观察了胶质纤维酸性蛋白（GFAP）、2',3'-环核苷酸磷酸二酯酶（CNPase）和神经丝（NF）的表达情况。当妊娠达到70%（15天）时，将大肠杆菌接种到孕鼠子宫角，对照组接种生理盐水。采用免疫组织化学法评估出生后第7天（P7）幼鼠脑中GFAP、CNPase和NF的表达，并用逆转录聚合酶链反应（RT-PCR）分析巨噬细胞炎性蛋白-1α mRNA（MIP-1α mRNA）、巨噬细胞炎性蛋白-1β mRNA（MIP-1β mRNA）、活化正常T细胞表达和分泌的趋化因子mRNA（RANTES mRNA）以及嗜酸性粒细胞趋化因子mRNA在出生后第1天（P1）、第3天（P3）和第7天（P7）幼鼠脑中的表达。与对照组相比，大肠杆菌处理组幼鼠在P7时脑室周围白质和海马中GFAP阳性细胞数量明显增加，但两组胼胝体中GFAP表达水平无显著差异。与对照组相比，大肠杆菌处理组幼鼠在P7时脑室周围白质和胼胝体中CNPase阳性染色的积分密度（ID）明显降低。与对照组相比，大肠杆菌处理组幼鼠在P7时脑室周围白质中NF阳性染色的ID明显降低，两组胼胝体中NF表达水平无显著差异。大肠杆菌处理的幼鼠在P1时脑中MIP-1α mRNA和MIP-1β mRNA的表达高于对照组，但在P3和P7时幼鼠脑中MIP-1α mRNA和MIP-1β mRNA的表达在两组间无显著差异。宫内感染后新生大鼠脑中GFAP、CNPase和NF表达的改变表明宫内感染可导致新生儿白质损伤。此外，宫内感染后新生儿脑中趋化因子如MIP-1α、MIP-1β表达的短暂增加表明MIP-1α、MIP-1β可能是介导新生儿白质损伤与宫内感染之间关系的一种机制。
