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目的研究小鼠脑组织同源盒基因(homeobox gene,Hox)的表达状态及人类巨细胞病毒(human cytomegalovirus,HCMV)感染对小鼠脑组织Hox基因表达的影响,以探讨HCMV致畸的分子机制。方法昆明系小鼠48只,随机分为实验对照组(16只)和病毒感染组(32只),病毒感染为脑内注射。在感染后7、15、30、60d分别以病理学方法观察病理损伤,以免疫组化方法检查HCMV-LA(late antigen),PCR检测小鼠脑组织中HCMV-DNA。在建立HCMV脑部感染的小鼠模型基础上,用RT-PCR测定Hox基因在病毒感染组和实验对照组小鼠脑组织中的表达,并用同位素标记的cDNA探针进行Northern-blot检测相应的表达状况。结果(1)接种HCMVAD169株的小鼠脑组织发生了病理改变,免疫组化和PCR方法在神经细胞内查到了HCMV-LA和DNA;(2)RT-PCR和Northern-blot发现:对照组的小鼠脑组织表达Hox-A9、Hox-A11、Hox-A12和Hox-A13,但不表达Hox-B13;HCMV感染后,小鼠脑组织被诱导表达Hox-B13,与对照组比较差异有统计学意义(P<0.01),且于感染后的30d达到高峰,而Hox-A9、Hox-A11的表达下调(P<0.05);Hox-A10和Hox-A13的表达增强。结论HCMVAD169株可感染小鼠神经系统。HCMV感染可诱导小鼠神经系统发育基因Hox表达改变,这对研究HCMV感染致畸的分子机制提供了有价值的理论依据。
Objective To investigate the effect of homeobox gene (Hox) expression in brain tissue of mice and the expression of Hox gene in mice brain tissue infected by human cytomegalovirus (HCMV) in order to explore the molecular mechanism of HCMV teratogenicity . Methods Forty-eight Kunming mice were randomly divided into experimental group (n = 16) and virus-infected group (n = 32). The virus infection was intracerebral injection. At 7, 15, 30 and 60 days after infection, the pathological changes were observed by pathology. HCMV-LA (late antigen) was detected by immunohistochemistry. HCMV-DNA was detected by PCR in mice brain. Based on the mouse model of HCMV infection in the brain, the expression of Hox gene was detected by RT-PCR in the brain tissue of mice infected with virus and the experimental control group. Northern blot was used to detect the corresponding Expression status. Results (1) HCMV-LA and DNA were detected in neurons of HCMVAD169-infected mice by immunohistochemistry and PCR. (2) RT-PCR and Northern-blot showed that in the control group Hox-A9, Hox-A11, Hox-A12 and Hox-A13 were not expressed in mouse brain tissue; Hox-B13 was induced in brain tissue of mice after HCMV infection, which was statistically different from the control group (P <0.01), and peaked at 30 days after infection, while the expression of Hox-A9 and Hox-A11 was down-regulated (P <0.05); the expression of Hox-A10 and Hox-A13 was enhanced. Conclusion HCMVAD169 can infect the nervous system of mice. HCMV infection can induce the change of neuronal development gene Hox expression in mice, which provides a valuable theoretical basis for studying the molecular mechanism of teratogenicity of HCMV infection.