Cell proliferation and nNOS expression in SH-SY5Y cells with co-cultivation of fluorosis and 7-nitroindazole

DENG Mingfen; ZHU Dan; GUI Chuanzhi; GUAN Zhizhong

doi:10.3969/j.issn.1002-266X.2017.11.007

Abstract：Objective To observe the cell proliferation and neuronal nitric oxide synthase (nNOS) expression in human neuroblastoma (SH-SY5Y) cells with co-cultivation of fluorosis and 7-nitroindazole.Methods SH-SY5Y cells cultured in vitro were divided into six groups, the control group, low-and high-dose fluoride exposure groups (0 mmol/L NaF, 0.2 mmol/L NaF and 2.0 mmol/L NaF), 7-NI group, low-and high-dose fluoride exposure combined with 7-NI groups (1 μmol/L 7-NI, 0.2 mmol/L NaF+1 μmol/L 7-NI , and 2.0 mmol/L NaF+1 μmol/L 7-NI), and cells in the above groups were treated for 24 h, and setting six holes in each group.The proliferation of SH-SY5Y cells was detected by CCK-8 (shown as OD450).The expression levels of nNOS mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blotting.Results Compared with the control group, low-and high-dose fluoride exposure groups exhibited significantly lower OD450 of SH-SY5Y cells (all P<0.05), while there was no statistically significant difference in 7-NI group (P>0.05).Compared with the low-dose fluoride group, the high-dose fluoride group exhibited significantly lower OD450 (P<0.05).Compared with the cells only treated with NaF, the OD450 of SH-SY5Y cells in the fluoride combined with 7-NI groups were significantly increased (all P<0.05).Compared with the control group, the expression levels of nNOS mRNA and protein in the low-and high-dose fluoride groups were significantly increased (all P<0.05), and the high-dose fluoride group was significantly higher than low-dose fluoride fluoride group (P<0.05).Compared with cells only treated with NaF, the expression levels of nNOS mRNA and protein in the fluoride combined with 7-NI groups were significantly decreased (all P<0.05).Conclusion Fluoride can inhibit the proliferation of SH-SY5Y cells, and co-cultivation of fluorosis and 7-nitroindazole can reduce the inhibitory effect of fluoride on the proliferation of SH-SY5Y cells.