Min Shi, Jian-bo Li, Hong-man Zhang, Ling-fei Yan, Min Xie, Jia-wei Chen
Nutr Neurosci. 2012 Nov;15(6):264-70. doi: 10.1179/1476830512Y.0000000022.
We aimed to explore neuritin expression in Schwann cells under different glucose conditions. Expression of neuritin at the levels of transcription and translation in purified Schwann cells was detected and measured using reverse transcriptase (RT) (quantitative) polymerase chain reaction (PCR) and western blot. Apoptosis of Schwann cells was measured by flow cytometry using Fluorescence Activated Cell Sorter (FACS) analysis and caspase fluorometric assay. Neuritin mRNA and protein were detected in cultured primary Schwann cells. Neuritin was identified as cell membrane form of protein and predominately as secreted or solube form of protein. Neuritin was significantly lower in 150 mM glucose condition, and more significantly lower in 300 mM glucose, than 5.6 mM glucose condition at 36 hours and especially at 48 hours of the culture, respectively (P < 0.05-0.01). In contrast to 5.6 mM glucose, obvious apoptosis of Schwann cells was demonstrated at 42 hours in 300 mM glucose condition and at 48 hours in 150 mM glucose, respectively (P < 0.05-0.01). Neuritin and apoptosis were correlated in a power regression (P < 0.01). 5.6 mM glucose cultured cells did not show these obvious changes during the experiment. It is concluded that neuritin mRNA and protein were expressed and down-regulated in Schwann cells under high-glucose concentration and the down-regulation may contribute to apopotosis of Schwann cells.
我们旨在探究不同葡萄糖条件下施万细胞中神经突蛋白的表达情况。使用逆转录(RT)(定量)聚合酶链反应(PCR)和蛋白质印迹法检测并测定纯化施万细胞中转录和翻译水平的神经突蛋白表达。通过使用荧光激活细胞分选仪(FACS)分析和半胱天冬酶荧光测定法的流式细胞术来检测施万细胞的凋亡。在培养的原代施万细胞中检测到神经突蛋白mRNA和蛋白质。神经突蛋白被鉴定为蛋白质的细胞膜形式,主要为分泌型或可溶性蛋白质形式。在培养36小时尤其是48小时时,150 mM葡萄糖条件下神经突蛋白明显低于5.6 mM葡萄糖条件,300 mM葡萄糖条件下则更低(P < 0.05 - 0.01)。与5.6 mM葡萄糖相比,300 mM葡萄糖条件下施万细胞在42小时时出现明显凋亡,150 mM葡萄糖条件下在48小时时出现明显凋亡(P < 0.05 - 0.01)。神经突蛋白与凋亡呈幂回归相关(P < 0.01)。5.6 mM葡萄糖培养的细胞在实验期间未显示出这些明显变化。结论是,在高葡萄糖浓度下施万细胞中表达并下调了神经突蛋白mRNA和蛋白质,这种下调可能导致施万细胞凋亡。