摘要
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进行测定 ,以探讨先天性HCMV感染致脑损害的机制。方法 在建立先天性HCMV中枢神经系统 (CNS)感染胎鼠模型的基础上 ,用逆转录 聚合酶链式反应 (RT PCR)测定受不同病毒剂量感染的胎鼠大脑皮层ET 1mRNA ,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转录量及胞内定位。结果 在大脑皮层组织的上清液中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脑炎性改变 ,并在神经细胞内发现特异性核内嗜碱性包涵体。RT PCR和原位杂交研究发现 ,受染胎鼠大脑皮层内ET 1mRNA转录量增加 ,以 1 0ml和 0 5ml组为显著 ,而 0 2 5ml组与正常对照组比较无明显差别。结论 HCMV可经胎盘垂直传播至胎鼠脑组织。先天性HCMV感染可刺激受染胎鼠CNSET 1mRNA的转录 ,且与母鼠所接种的病毒量存在一定的量效关系。这些结果提示 ,ET 1在先天性HCMV感染脑损害过程中 ,早期可导致组织缺血性改变 ,而晚期则与受损大脑皮层的功能恢复有关。这对了解先天性HCMV感染致CNS损伤的机理将提供有价值的参考依据 。
Objective To explore mechanisms of brain damage following congenitally infected human cytomegalovirus, the transcription of endothelin 1 (ET 1) mRNA of fetal mouse cerebral cortex (HCMV) were analyzed. Methods On the basis of developing congenital HCMV infective fetal model, reverse transcriptase polymerase chain reaction (RT PCR) was used to determine ET 1 mRNA of fetal mouse cerebral cortex infected by different inoculum size meanwhile the intracellular location of mRNA's was conducted with in situ hybridization by digoxigenin labelled ET 1 mRNA oligonucleotide probe. Virus isolation and sections coated with histostik applied to stained with hematoxylin and eosin (HE) were synchronously carried out. Results HCMV were isolated from the tissue supernatant. Histopathologic examination show that pathological changes consisted of destructive meningoencephalitis and large, basophilic, intranuclear neuronal inclusions of virus in fetal mouse cerebral cortex. RT PCR and in situ hybridization showed that ET 1 mRNA in cerebral cortex of fetal mouse increased significantly. The results from 1.0ml and 0.5ml group are significant, while the result of 0.25ml group has no significant difference no matter whether pathological change or the content of ET 1 mRNA compared with normal control group. Conclusions Our results suggest that HCMV is able to cause fetal mouse CNS infection as a result of transplacental transmission. Congenital HCMV infection stimulated the enhanced transcription of ET 1 mRNA of fetal mouse cerebral cortex in a dose dependent manner. ET 1, functions as a neuroregulator, may have a dual effects on the brain damage caused by congenital HCMV infection. Those provides a rational for understanding the mechanism of CNS pathogenesis by congenital HCMV infection and provides a valuable method for clinical prevention and cure at the same time.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2001年第1期25-29,共5页
Chinese Journal of Microbiology and Immunology
基金
安徽省自然科学基金资助项目! (95 医 18)
