文章摘要
郭丹丹,姚 堃,周 锋,李凌云,茌 静,王 芳,顾 斌,张 春.人类疱疹病毒6A对神经细胞的嗜性及细胞周期的影响[J].南京医科大学学报,2011,(7):970~975
人类疱疹病毒6A对神经细胞的嗜性及细胞周期的影响
Study on neural cell tropism of human herpesvirus 6 and its effect on neural cell cycle
投稿时间:2010-12-14  
DOI:10.7655
中文关键词: 人类疱疹病毒6A  神经细胞  细胞嗜性  细胞周期
英文关键词: human herpesvirus 6A  neural cells  cell tropism  cell cycle
基金项目:国家自然科学基金(30972784);南京医科大学科技发展基金重点项目(08NMUZ003)
作者单位
郭丹丹 南京医科大学微生物学与免疫学系,江苏 南京 210029 
姚 堃 南京医科大学微生物学与免疫学系,江苏 南京 210029 
周 锋 南京医科大学微生物学与免疫学系,江苏 南京 210029 
李凌云 南京医科大学微生物学与免疫学系,江苏 南京 210029 
茌 静 南京医科大学微生物学与免疫学系,江苏 南京 210029 
王 芳 南京医科大学微生物学与免疫学系,江苏 南京 210029 
顾 斌 南京医科大学微生物学与免疫学系,江苏 南京 210029 
张 春 南京医科大学微生物学与免疫学系,江苏 南京 210029 
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中文摘要:
      目的:研究人类疱疹病毒6A对神经细胞嗜性及细胞周期的影响?方法:HHV-6A GS株感染神经细胞,倒置显微镜观察细胞病变?PCR法鉴定神经细胞中HHV-6A U22基因,实时荧光定量PCR法测定神经细胞中HHV-6A U22基因的相对含量?免疫细胞化学和Western blot检测神经细胞中HHV-6A糖蛋白gB的表达?神经细胞感染HHV-6A后MTT法测定细胞增殖的改变,流式细胞仪检测神经细胞周期的改变?结果:HHV-6A GS株感染5天后,U373细胞形态无明显变化,SK-N-SH和SHG44细胞聚集,折光性降低,细胞发生明显病变?PCR法检测到U373?SK-N-SH和SHG44细胞中均含有HHV-6A U22基因,实时荧光定量PCR法发现随着感染天数的增加,HHV-6A DNA在细胞中含量逐渐降低?免疫细胞化学和Western blot结果显示病毒糖蛋白gB在细胞中表达,其中在U373细胞和SHG44细胞中的表达量高于SK-N-SH细胞?MTT法显示HHV-6A促进U373细胞和SHG44细胞的增殖,抑制SK-N-SH细胞的增殖?流式细胞仪检测细胞周期,结果显示U373和SHG44细胞感染组与对照组相比,G1期细胞百分数减少,S和G2期细胞百分数增多;SK-N-SH细胞G1期细胞百分数增多,而S和G2期细胞百分数减少?结论:HHV-6A感染神经细胞,对神经胶质细胞的嗜性强于神经元细胞?HHV-6A通过改变神经细胞周期进程,促进神经胶质细胞增殖,而抑制神经元细胞增殖?
英文摘要:
      Objective:To study neural cell tropism of human herpesvirus 6A(HHV-6A) and its effect on neural cell cycle. Methods:The cell morphology was assessed by a light microscope. The fragment of HHV-6A U22 gene was amplified by PCR and relative amount of HHV-6A U22 gene was analyzed using quantitative real-time PCR. Expression of HHV-6A gB was examined by immunocytochemical and Western blot analysis. Cell proliferation was measured by MTT assay. Cell cycle analysis was evaluated by the propidium iodide assay. Results:On the fifth day after being infected by HHV-6A,U373 cells had no obvious change,but SK-N-SH and SHG44 cells showed cytopathic effect. HHV-6A U22 gene was detected in all three infected cell lines and its relative amounts kept reducing all the time. HHV-6A gB was positive in the three infected cell lines,furthermore, the expression levels in U373 and SHG44 were more than those in SK-N-SH. HHV-6A stimulated the proliferations of U373 and SHG44 cells,however,inhibited the proliferation of SK-N-SH cells significantly. Compared with uninfected cells,percentages of U373 and SGH44 cells were decreased in phase G1 but increased in phase S and G2 after cells were infected by HHV-6A,as to SK-N-SH,the percentages were increased in phase G1 but decreased in S and G2 phases. Conclusion:All three neural cell lines can sustain replication of HHV-6A,and moreover,HHV-6A can boost astrocyte proliferation and inhibit neuronal cell proliferation.
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