巨细胞病毒感染后新生小鼠脑内Wif-1、Shh的表达变化

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目的:探讨鼠巨细胞病毒(murine cytomegalovirus,MCMV)感染后新生小鼠脑内Wnt inhibitory factor-1(Wif-1)、Sonic hedgehog(Shh)信号分子的表达变化及对中枢神经系统发育的可能影响。方法:体外3T3细胞培养、MCMV接种增殖;60只生后3 d的BALB/c仔鼠随机分为正常对照组及病毒感染组,病毒组通过腹腔注射法建立新生小鼠中枢神经系统MCMV感染模型,而对照组则注射等量的生理盐水;每组设立3个时间点亚组(造模后3 d,7 d,14 d),相应时点处死仔鼠,取脑组织备用。通过RT-PCR检测两组各时点脑组织中的MCMV-DNA表达;Western Blot检测小鼠脑组织中Wif-1、Shh蛋白的表达;流式细胞术检测两组的细胞周期比例改变。结果:病毒组小鼠脑组织中MCMV-DNA的表达随着时间推移持续增多,而对照组3个时间点均阴性表达;病毒组小鼠脑组织中Wif-1分子的表达相对于对照组增强,而Shh分子的表达相比于对照组则表现出明显的抑制;病毒组小鼠脑组织中处于分裂期的细胞百分比较对照组下降。结论:MCMV感染后的新生小鼠脑组织内Wif-1表达增强,而Shh信号分子则下调,其表达异常变化可能影响Wnt、Hedgehog信号通路,并影响中枢神经系统成熟过程。 OBJECTIVE: To investigate the changes of Wnt inhibitory factor-1 (Wif-1) and Sonic hedgehog (Shh) signaling molecules in brain of murine cytomegalovirus (MCMV) infection and their possible impact on central nervous system development . Methods: 3T3 cells were cultured in vitro and MCMV was inoculated to proliferate. Sixty BALB / c mice aged 3 days after birth were randomly divided into normal control group and virus-infected group. The model group was established by intraperitoneal injection to establish MCMV infection model of neonatal mouse central nervous system , While the control group was injected with the same amount of saline. Each group was divided into 3 sub-groups (3 days, 7 days and 14 days after modeling). The pups were sacrificed at the corresponding time points, and brain tissue was taken for later use. The expression of MCFV-DNA in brain tissue was detected by RT-PCR at each time point. The expression of Wif-1 and Shh protein in brain tissue was detected by Western Blot. The proportion of cell cycle was detected by flow cytometry. Results: The expression of MCMV-DNA in the brain tissue of the virus-infected mice continued to increase over time, while the control group was negatively expressed at all three time points. The expression of Wif-1 in the brain tissue of the virus-infected mice was increased compared with that of the control group , While the expression of Shh was significantly inhibited compared with the control group; the percentage of cells in the dividing stage in the brain tissue of the virus group was lower than that in the control group. CONCLUSION: Wif-1 expression is up-regulated in brain tissue of MCMV-infected neonatal mice, whereas Shh signaling is down-regulated. The abnormal expression of Wif-1 may affect the Wnt and Hedgehog signaling pathway and affect the central nervous system maturation process.
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