吡咯喹啉醌对大鼠雪旺细胞形态及增殖细胞核抗原表达的影响

Effect of Pyrroloquinoline Quinone on Morphology of Cultured Rat Schwann Cells and the Expression of Proliferating Cell Nuclear Antigen

目的:观察吡咯喹啉醌(PQQ)对体外培养雪旺细胞的形态、细胞数量及雪旺细胞内增殖细胞核抗原(PCNA)表达的影响。方法:从出生3-5dSD大鼠坐骨神经分离培养雪旺细胞,纯化并经S-100免疫荧光标记鉴定;设立对照组和实验组,分别加入100nmol/LPQQ和等量PBS培养48h后倒置显微镜下观察细胞形态并进行细胞计数;将不同浓度的PQQ(0,10,50,100,500nmol/L)加入雪旺细胞培养基中,培养48h后提取总mRNA及总蛋白进行PCNA的RT-PCR和Western blotting检测。结果:100nmol/LPQQ干预雪旺细胞48h可以使细胞的形态发生变化并使细胞数量增加,细胞数量较对照组增加13.6%;RT-PCR及Western blotting检测结果表明:PQQ浓度在10-500nmol/L浓度范围内可增加雪旺细胞内PCNA的表达,且当PQQ浓度为100nmol/L时该上调效应最明显,分别是对照组的2.0倍和3.4倍。结论:PQQ可增加体外培养雪旺细胞的数量;并可使体外培养雪旺细胞内PCNA表达上调。

Objective：To investigate the effects of pyrroloquinoline quinine （PQQ） on the morphology and proliferation of rat Schwann cells, and on the expression of proliferating cell nuclear antigen （PCNA） in cultured rat Schwann cells. Methods： Schwann cells were obtained and cultured from sciatic nerves of 3-5 day SD rats in vitro, then were identified and purified by immunofluorescence against S-100. Cells of experimental group were treated with 100 nmol/L PQQ, control group were treated with PBS, and both groups were cultured for 48 hours to observe the morphology of Schwann cells and to count the cell number by inverted microscope. The expression of PCNA in Schwann cells treated with PQQ at different concentrations （0, 10, 50, 100, and 500 nmol/L） was determined by RT-PCR and Western blot. Results： PQQ with a concentration of 100 nmol/L could affect the cell morphology and increase the cell number （1.36 times, versus control）. PQQ with concentrations ranged 10-500 nmol/L could increase the expression of PCNA mRNA and protein, and the PCNA expression reached the peak in 100 nmol/L PQQ treated Schwann cells （mRNA 2.0 times and protein 3.4 times, respectively, versus control）. Conclusion： PQQ could increase the cell number of cultured Schwann cells, and up-regulate the expression of PCNA in cultured Schwann cells.