模拟微重力及病毒作用下神经细胞生物活性变化规律研究  

Study on the Biological Activity of Nerve Cells under the Action of Virus and Simulated Microgravity

作　　者：裴思竹 孙飞一[1] 马宏[1] PEI Sizhu;SUN Feiyi;MA Hong(School of Life Science,Beijing Institute of Technology,Beijing,100081)

机构地区：[1]北京理工大学生命学院,北京100081

出　　处：《生命科学仪器》2019年第6期60-64,共5页Life Science Instruments

基　　金：国家自然基金面上项目,项目编号:8177071105。

摘　　要：空间环境会对人的骨骼、神经系统、免疫系统等产生不利影响,其中微重力条件是重要的影响因素。研究表明微重力条件下重组腺相关病毒(AAV)的活性会增加,也有研究表明病毒的活性变化和自噬有一定的关系。首先构建重组AAV病毒,并与增强型绿色荧光蛋白(eGFP)基因一起包装,并在微重力条件下感染U87神经胶质细胞和SH-SY5Y神经元;使用荧光显微镜和流式细胞仪等技术检测结果。经研究,微重力能促进AAV转导入细胞并促进基因表达,影响细胞的粘附能力,并能影响病毒感染受体的变化。为了检测自噬水平的变化,利用慢病毒构建了稳转株,通过荧光检测发现,微重力条件下自噬水平会增加。QPCR分析发现微重力下,细胞中内体增多,从而促进病毒的转运。这些发现有助于了解微重力对rAAV侵染神经细胞的影响,对基因治疗和在空间环境下研究病毒神经致病性的机制有重要意义。Space environment will have adverse effects on human bones,nervous system and immune system,among which microgravity condition is an important factor.Studies have shown that the activity of recombinant adeno-associated virus(AAV)increases under microgravity,and some studies have also shown that the activity of the virus is related to autophagy.First,the recombinant AAV virus was constructed and packaged with the enhanced green fluorescent protein(eGFP)gene,and infected with U87 glioma cells and SH-SY5Y neurons under microgravity.Fluorescence microscopy and flow cytometry were used to detect the results.Microgravity can promote the transfer of AAV into cells and promote gene expression,affect cell adhesion,and affect the changes of virus infection receptors.In order to detect the changes in autophagy level,a stable strain was constructed using lentivirus.Fluorescence detection showed that the autophagy level would increase under microgravity.QPCR analysis showed that microgravity increased the number of endosomes in cells,which promoted the transport of virus.These findings are helpful for understanding the effect of microgravity on rAAV infected nerve cells,and are of great significance for gene therapy and the study of the mechanism of viral neuro pathogenicity in space environment.

关 键 词：模拟微重力 腺相关病毒 神经细胞 

分 类 号：Q2[生物学—细胞生物学]