未经证实:本研究旨在了解缺氧暴露后人类海马星形胶质细胞的特征。根据初步筛选,选择15分钟作为时间点,并将细胞暴露于不同的氧百分比。
未经鉴定:用于检查细胞死亡的台盼蓝活力测定。免疫荧光分析,胶质纤维酸性蛋白(GFAP)用于描绘星形胶质细胞的形态。进行缺氧诱导因子1(HIF-1)染色以确认缺氧诱导的细胞死亡,并且与对照相比,暴露的星形胶质细胞中HIF-1α的显著表达。在分子水平上,选择了基因,如甘油醛3-磷酸脱氢酶(GAPDH),GFAP,HIF-1α和B细胞淋巴瘤2(Bcl-2),并进行逆转录聚合酶链反应(RT-PCR)。
UNASSIGNED:显微镜在对照中显示出丝状和透明的细胞核外观,而在3%的氧气中发现了没有细胞刚性结构的破裂核。对照和缺氧细胞也用膜联蛋白V-异硫氰酸荧光素(膜联蛋白V-FITC)染色。荧光显微镜下显示星形胶质细胞缺氧后显示出较高的细胞核表达,而对照组则没有。PI和FITC的合并显示了对照和缺氧之间细胞核表达的差异。在分子分析中,GFAP有显著变化,与对照组相比,缺氧暴露细胞中的HIF-1α和Bcl-2。
未经鉴定:暴露于缺氧(3%氧气15分钟)的细胞明显显示出损伤。获得了人海马星形胶质细胞基因组对缺氧反应的一般看法。
UNASSIGNED: The present study aimed to understand the characterisation of human hippocampal astrocyte following hypoxia exposure. Based on the preliminary screening, 15 min was chosen as the time point and the cells were exposed to different oxygen percentages.
UNASSIGNED: The Trypan blue viability assay used to examine cell death. Immunofluorescence assay, glial fibrillary acidic protein (GFAP) was used to portray the morphology of astrocytes. The hypoxia-inducible factor 1 (HIF-1) staining was performed to confirm hypoxia induced cell death and there was a dramatic expression of HIF-1α displayed in exposed astrocyte cells compared to the control. In molecular level, genes were chosen, such as glyceraldehyde 3-phosphate dehydrogenase (GAPDH), GFAP, HIF-1α and B-cell lymphoma 2 (Bcl-2) and ran the reverse transcription-polymerase chain reaction (RT-PCR).
UNASSIGNED: Microscope revealed a filamentous and clear nucleus appearance in a control whereas the rupture nuclei with no rigid structure of the cell were found in the 3% oxygen. The control and hypoxia cells were also stained with the annexin V-fluorescein isothiocyanate (annexin V-FITC). Fluorescence microscope reveals astrocyte cells after hypoxia showed higher expression of nuclei but not in control. Merging PI and FITC showed the differences of nuclei expression between the control and hypoxia. In the molecular analysis, there were significant changes of GFAP, HIF-1α and Bcl-2 in hypoxia exposed cells when compared to the control group.
UNASSIGNED: Cells that were exposed to hypoxia (3% oxygen for 15 min) clearly showed damage. General view of human hippocampal astrocyte genomic response to hypoxia was obtained.