CAJ | 학술논문

目的建立鼠巨细胞病毒(MCMV)播散性感染的小鼠模型,在整体水平观察脾脏病理损伤与促炎因子IL-17A表达的关系,初步探讨IL-17A参与巨细胞病毒感染后脾脏病理改变的机制.方法建立鼠巨细胞病毒全身播散性感染模型,MCMV Smith株腹腔接种后3、7、14、28 d各处死小鼠3只,同时设正常小鼠作为模拟感染对照.标准空斑实验检测脾脏的病毒滴度,RT-PCR法测定脾脏中MCMV mRNA、IL-17A mRNA的表达,免疫组化法检测脾脏的IL-17A蛋白的表达,HE染色法评估脾脏的病理性损伤程度,分析IL-17A的表达与脾脏病理改变的关系.结果脾组织的病毒滴度在巨细胞病毒感染后3d升高,7d有所降低,在感染后第14天用标准空斑实验检测不到病毒.MCMV mRNA在病毒感染后3、7d可以检测的到；IL-17A mRNA的表达,与模拟感染对照组相比,MCMV感染组表达逐渐增加,并在14 d达到高峰,28 d显著下降.免疫组化法检测的脾脏IL-17A蛋白的表达也在14 d达到高峰.脾组织的病理损伤逐渐加重,至感染后第14天病变达高峰后逐渐减轻.脾脏组织病理损伤最重与IL-17A高表达出现在同一时期.结论促炎因子IL-17A的高表达与脾脏组织的病理损伤程度呈现明显相关性.
목적 건립서거세포병독(MCMV)파산성감염적소서모형,재정체수평관찰비장병리손상여촉염인자IL-17A표체적관계,초보탐토IL-17A삼여거세포병독감염후비장병리개변적궤제.방법 건립서거세포병독전신파산성감염모형,MCMV Smith주복강접충후3、7、14、28 d각처사소서3지,동시설정상소서작위모의감염대조.표준공반실험검측비장적병독적도,RT-PCR법측정비장중MCMV mRNA、IL-17A mRNA적표체,면역조화법검측비장적IL-17A단백적표체,HE염색법평고비장적병이성손상정도,분석IL-17A적표체여비장병리개변적관계.결과 비조직적병독적도재거세포병독감염후3d승고,7d유소강저,재감염후제14천용표준공반실험검측불도병독.MCMV mRNA재병독감염후3、7d가이검측적도；IL-17A mRNA적표체,여모의감염대조조상비,MCMV감염조표체축점증가,병재14 d체도고봉,28 d현저하강.면역조화법검측적비장IL-17A단백적표체야재14 d체도고봉.비조직적병리손상축점가중,지감염후제14천병변체고봉후축점감경.비장조직병리손상최중여IL-17A고표체출현재동일시기.결론 촉염인자IL-17A적고표체여비장조직적병리손상정도정현명현상관성.
Objective To explore the correlation between the expression of IL-17A and the degree of spleen damage in acute mouse cytomegalovirus(MCMV) disseminated infection in vivo and to understand the mechanism about how IL-17A involved in the pathological damage of the spleen in MCMV infection.Methods An acute disseminated MCMV infection model was established in mice.BALB/c mice were randomly divided into two groups.Mice in group one were infected with MCMV Smith to establish disseminated infection.Mice in another group were sham-infected control.Three mice from each group were randomly chosen to be sacrificed on days 3,7,14 and 28 after the infection.Viral titers in spleen tissues were determined using a standard plaque assay.The expression of IL-17A mRNA and MCMV mRNA in the splenocytes were measured by RT-PCR.The expression of IL-17A in spleen tissues was observed by immunohistochemical staining.The pathology of the infected mice was assessed by histological examination of H&E stained spleen sections.Results Viral titers and MCMV mRNA in the spleen peaked on day 3,but quickly diminished on day 7.Virus was no longer detectable in the spleen on day 14 after the infection.The expression of IL-17A mRNA was significantly increased during the acute infection and reached the highest level on day 14,then decreased on day 28.It is significantly higher than that of the mock infection group.Immunohistochemistry assay also indicated that the expression of IL-17A in spleen tissue gradually increased to climax on day 14,then decreased on day 28.Accordingly,the pathological damages of spleen tissue in the infected mice deteriorated until day 14,then showed signs of recovery on day 28.The most severe pathological injury of spleen tissue and the highest expression of IL-17A appeared in the same period of time.Conclusion Our results showed a close correlation between IL-17A and the pathological damage in spleen.Thus,IL-17A may contribute to the spleen pathological damage during the acute disseminated MCMV infection.