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目的探讨全氟辛烷磺酸(PFOS)对大鼠神经干细胞增殖和迁移的影响及其机制。方法分离怀孕15 d(E15)SD大鼠大脑皮质,神经上皮干细胞蛋白染色鉴定培养的神经干细胞。PFOS 10,50和100μmo·lL-1处理大鼠神经干细胞24 h后,Edu免疫荧光染色检测神经干细胞的增殖;荧光定量聚合酶链反应法(qPCR)检测样本中Stat3,Sox2和Notch1 mRNA表达;Western蛋白质印迹法检测Stat3及p-Stat3蛋白表达;采用Transwell小室检测神经干细胞的迁移能力。结果经鉴定分离培养的原代细胞为神经干细胞。Edu免疫荧光染色结果显示,与正常对照组相比,经PFOS处理的神经干细胞的Edu阳性率明显下降(P<0.05),说明PFOS抑制神经干细胞的增殖;qPCR检测结果显示,PFOS处理组神经干细胞中Stat3 mRNA表达显著降低(P<0.05);Western蛋白质印迹实验结果表明,PFOS组p-Stat3蛋白的表达降低(P<0.05),但总Stat3蛋白表达无变化;Transwell小室检测结果显示,PFOS具有抑制神经干细胞迁移的能力(P<0.05)。结论 PFOS可导致神经干细胞增殖和迁移能力降低,可能与抑制Stat3的转录和翻译后的磷酸化过程有关。
Objective To investigate the effect of perfluorooctane sulfonate (PFOS) on the proliferation and migration of neural stem cells in rats and its mechanism. Methods Cultured neural stem cells were isolated from the cerebral cortex of 15 d (E15) SD rats and the protein staining of neural epithelial stem cells. After treated with PFOS at 10, 50 and 100μmol · L-1 for 24 h, the proliferation of neural stem cells was detected by Edu immunofluorescence staining; the expression of Stat3, Sox2 and Notch1 mRNA was detected by qPCR; Western blotting was used to detect the expression of Stat3 and p-Stat3 protein. Transwell chamber was used to detect the migration of neural stem cells. Results The primary cells isolated and identified were neural stem cells. Edu immunofluorescence staining showed that compared with the normal control group, the Edu positive rate of PFOS-treated neural stem cells was significantly decreased (P <0.05), indicating that PFOS inhibited the proliferation of neural stem cells; qPCR test results showed that PFOS-treated neural stem cells (P <0.05). Western blotting results showed that the expression of p-Stat3 protein in PFOS group was decreased (P <0.05), but the expression of Stat3 mRNA was not changed. Transwell chamber assay showed that PFOS had Inhibit the migration of neural stem cells (P <0.05). Conclusions PFOS can reduce the proliferation and migration of neural stem cells, which may be related to the inhibition of Stat3 transcription and posttranslational phosphorylation.