氟及7-NI共培养的SH-SY5Y细胞nNOS表达、增殖情况观察

发布时间：2018-05-02 06:31

本文选题：氟 + 氟中毒　；参考：《山东医药》2017年11期

【摘要】：目的观察氟及神经型一氧化氮合成酶(nNOS)特异性抑制剂7-硝基吲唑(7-NI)共培养的SH-SY5Y细胞增殖情况和nNOS表达变化。方法将体外培养SH-SY5Y细胞分为对照组、低氟组、高氟组、7-NI组、低氟+7-NI组、高氟+7-NI组,每组6个复孔。低氟组加入0.2 mmol/L Na F,高氟组加入2.0 mmol/L Na F,7-NI组加入1μmol/L 7-NI,低氟+7-NI组加入0.2 mmol/L Na F和1μmol/L 7-NI,高氟+7-NI组加入2.0 mmol/L Na F和1μmol/L 7-NI,对照组加入等体积培养液,继续培养24 h。采用CCK-8法检测各组SH-SY5Y细胞增殖情况(以OD450表示);采用实时荧光定量PCR法检测各组SH-SY5Y细胞NOS mRNA;采用Western blotting法检测各组SH-SY5Y细胞NOS蛋白。结果低氟组、高氟组细胞OD450均低于对照组(P均0.05),且高氟组细胞OD450低于低氟组(P0.05);7-NI组细胞OD450与对照组相比P0.05;低氟+7-NI组细胞OD450高于低氟组(P0.05);高氟+7-NI组细胞OD450高于高氟组(P0.05)。低氟组、高氟组细胞nNOS mRNA和蛋白相对表达量均高于对照组(P均0.05),且高氟组细胞nNOS mRNA和蛋白相对表达量均高于低氟组(P均0.05);7-NI组细胞nNOS mRNA和蛋白相对表达量均低于对照组(P均0.05);低氟+7-NI组细胞nNOS mRNA和蛋白相对表达量均低于低氟组(P0.05);高氟+7-NI组细胞nNOS mRNA和蛋白相对表达量均低于高氟组(P0.05)。结论氟可以抑制SHSY5Y细胞增殖,7-NI共培养可以减轻氟对SH-SY5Y细胞的增殖抑制作用。
[Abstract]:Objective to observe the proliferation and nNOS expression of SH-SY5Y cells co-cultured with fluorine and neuronal nitric oxide synthase (NNOS) specific inhibitor 7-nitroindazolium. Methods SH-SY5Y cells cultured in vitro were divided into control group, low fluorine group, high fluorine group, high fluorine group, low fluorine 7-NI group and high fluorine 7-NI group. The low fluorine group added 0.2 mmol/L Na F, the high fluorine group added 2.0 mmol/L Na F 7 NI, the low fluorine 7-NI group added 0.2 mmol/L Na F and 1 渭 mol/L 7-NI, the high fluorine 7-NI group added 2.0 mmol/L Na F and 1 渭 mol/L 7-NI, and the control group added the same volume culture medium for 24 h. The proliferation of SH-SY5Y cells in each group was detected by CCK-8 assay (expressed as OD450), NOS mRNAs of SH-SY5Y cells in each group were detected by real-time fluorescence quantitative PCR assay, NOS protein of SH-SY5Y cells in each group was detected by Western blotting assay. Results the cell OD450 of low fluorine group and high fluoride group were lower than that of control group (P 0.05), and the OD450 of high fluorine group was lower than that of low fluorine group (P 0.05), the cell OD450 of high fluorine group was lower than that of control group (P 0.05), the cell OD450 of low fluorine 7-NI group was higher than that of low fluorine group (P 0.05), and the cell OD450 of high fluorine 7-NI group was higher than that of high fluoride group (P 0.05). Low fluorine group, The relative expression of nNOS mRNA and protein in the high fluorine group was higher than that in the control group (P 0.05), and the relative expression of nNOS mRNA and protein in the high fluoride group was higher than that in the low fluorine group. The relative expression of nNOS mRNA and protein in the high fluorine group was lower than that in the control group. The relative expression of nNOS mRNA and protein in the low fluorine 7-NI group was lower than that in the low fluorine group, and the relative expression of nNOS mRNA and protein in the high fluorine 7-NI group was lower than that in the high fluoride group. Conclusion fluoride can inhibit the proliferation of SHSY5Y cells and can reduce the inhibitory effect of fluoride on the proliferation of SH-SY5Y cells.
【作者单位】：贵州医科大学病理学教研室;贵州省肿瘤医院;贵州省分子生物学重点实验室;
【基金】：国家自然科学基金资助项目(81460482) 教育部博士学科点专项科研基金资助项目(20115215120003)
【分类号】：R595

【相似文献】