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目的 对先天性人巨细胞病毒 (HCMV)感染的胎鼠大脑皮层内皮素 1(ET 1)mRNA进行定性及定量分析 ,探讨先天性HCMV感染致脑损害的机制 ,为临床防治提供一种有价值的手段。方法 在建立HCMV先天性中枢神经系统 (CNS)感染胎鼠模型的基础上 ,用逆转录 聚合酶链反应 (RT PCR)测定受不同病毒剂量感染的胎鼠大脑皮质ET 1mR NA ,并用地高辛标记的ET 1寡核苷酸探针对大脑皮层细胞印片进行原位杂交以检测相应mRNA及胞内定位 ,并同步进行病毒分离和病理学检查。结果 病毒分离结果显示 ,在脑组织的上清液中HCMV分离阳性 ;病理学研究证实受染胎鼠大脑皮层表现为侵袭性脑膜炎性改变 ,并在神经细胞内发现特征性核内嗜碱性包涵体 ;受染胎鼠大脑皮层内ET mRNA增加 ,以 1 0ml和 0 5ml组为显著 (P<0 0 0 1) ,而 0 2 5ml组与正常对照组比较差别无显著性。原位杂交结果证实 ,ET 1mRNA主要存在于大脑皮层的胶质细胞中。结论 HCMV可经小鼠胎盘垂直传播至胎鼠脑组织。受染胎鼠大脑皮层ET 1mRNA明显增加 ,且这种结果与所接种的病毒量存在量效关系。ET 1mRNA主要存在于大脑皮层的胶质细胞中可能表明ET 1可作为一种神经调节肽以旁分泌和自分泌方式影响神经组织的功能 ,参与HCMV先天性感染脑损害的过程
Objective To investigate the mechanism of brain damage induced by congenital HCMV infection in congenital human cytomegalovirus (HCMV) infection by detecting the expression of endothelin-1 (ET 1) mRNA in fetal rat cortex, and to provide valuable information for clinical prevention and treatment s method. Methods The embryonic rat model of HCMV congenital central nervous system (CNS) infection was established. ET 1mR NA of fetal cerebral cortex was detected by reverse transcription-polymerase chain reaction (RT-PCR) Labeled ET 1 oligonucleotide probes were used to perform in situ hybridization of cortical cell prints to detect the corresponding mRNA and intracellular localization and simultaneous virus isolation and pathology. Results The result of virus isolation showed that the HCMV was separated in the supernatant of brain tissue. The pathological study confirmed that the infected cerebral cortex of the fetus showed invasive meningitis, and found characteristic intrahepatic basophilic The content of ET mRNA in the cerebral cortex of fetus fetus increased significantly in 10 ml and 0 5 ml groups (P <0.01), while there was no significant difference between 0 2 5 ml group and normal control group. In situ hybridization results confirmed that ET 1 mRNA is mainly present in the glial cells of the cerebral cortex. Conclusion HCMV can be transmitted perpendicularly to fetal rat brain tissue via mouse placenta. The ET 1 mRNA in the cerebral cortex of infected fetus significantly increased, and this result was correlated with the amount of virus inoculated. The predominant presence of ET 1 mRNA in glial cells of the cerebral cortex may indicate that ET 1 acts as a neurotrophin in a paracrine and autocrine manner affecting the function of neural tissue and is involved in the process of congenital infection of HCMV with brain damage