氟及7-NI共培养的SH-SY5Y细胞nNOS表达、增殖情况观察  

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

作　　者：邓明芬[1] 朱丹[1] 桂传枝[1,2] 官志忠[1,3] 

机构地区：[1]贵州医科大学病理学教研室,贵阳550004 [2]贵州省肿瘤医院 [3]贵州省分子生物学重点实验室

出　　处：《山东医药》2017年第11期22-25,共4页Shandong Medical Journal

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)

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(P<0.05);7-NI组细胞OD450与对照组相比P>0.05;低氟+7-NI组细胞OD450高于低氟组(P<0.05);高氟+7-NI组细胞OD450高于高氟组(P<0.05)。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(P<0.05);高氟+7-NI组细胞nNOS mRNA和蛋白相对表达量均低于高氟组(P<0.05)。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。Objective To observe the cell proliferation and neuronal nitric oxide synthase （nNOS） expression in hu- man neuroblastoma （SH-SY5Y） cells with co-cultivation of fluorosis and 7-nitroindazole. Methods SH-SY5Y cells cul- tured in vitro were divided into six groups, the control group, low-and high-dose fluoride exposure groups （0 mmol/L NaF, 012 mmol/L NaF and 2.0 mmol/L NaF） , 7-NI group, low-and high-dose fluoride exposure combined with 7-NI groups （1μmol/L 7-NI, 0.2 mmot/L NaF ＋ 1 μ mol/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-SYSY 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 exhibi- ted significantly lower OD450 of SH-SYSY cells （ all P 〈 0.05 ）, while there was no statistically significant difference in 7-NI group （P 〉 O. 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-SYSY 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 sig- nificantly decreased （ all P 〈 0.05 ）. Conclusion Fluoride can inhibit the proliferation of SH-SY5Y cells, and co-cultiva- tion of fluorosis and 7-nitroindazole can reduce the inhibitory effect of fluori

关 键 词：氟 氟中毒 人神经母细胞瘤细胞 7-硝基吲唑 神经型一氧化氮合成酶 一氧化氮 

分 类 号：R995[医药卫生—毒理学] R977.3[医药卫生—药学]