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目的探讨氟对原代大鼠海马细胞生长发育、神经细胞黏附分子(NCAM)mRNA 及蛋白表达的影响。方法原代培养大鼠海马细胞暴露于20、40和80μg/ml 氟化钠24 h 后,观察染氟前后海马细胞生长情况,采用噻唑蓝比色试验、逆转录-聚合酶链反应和蛋白印迹技术分别检测大鼠海马细胞存活情况、NCAM mRNA 和蛋白表达水平。结果与对照组相比,80μg/ml 剂量组海马细胞数量减少,突起减少、变短,细胞存活率下降。40、80μg/ml 剂量组 NCAM mRNA 水平均显著低于对照组,20μg/ml 剂量组 NCAM mRNA 水平虽与对照组之间差异无统计学意义,但存在下降的趋势,NCAM mRNA 水平与氟浓度呈现一定的剂量效应关系,且随染氟浓度升高而降低。40、80μmg/ml 剂量组细胞 NCAM-180蛋白表达水平显著低于对照组,各染氟组细胞 NCAM-140蛋白表达水平显著低于对照组,80μg/ml 剂量组细胞 NCAM-120蛋白表达水平显著低于对照组。结论氟可抑制海马细胞生长和存活,并使 NCAM mRNA 和蛋白表达水平下降,氟对发育中海马损伤可能是其神经毒性的作用位点之一。
Objective To investigate the effect of fluoride on the growth and expression of hippocampal cells and the expression of neural cell adhesion molecule (NCAM) mRNA and protein in primary rats. Methods Primary cultured rat hippocampal cells were exposed to 20, 40 and 80 μg / ml sodium fluoride for 24 h. The growth of hippocampal cells before and after fluoridation were observed. Thiazolyl blue colorimetric assay, reverse transcription - polymerase chain reaction and Western blot Technology were detected hippocampal cell survival, NCAM mRNA and protein expression levels. Results Compared with the control group, the number of hippocampal neurons in 80μg / ml group decreased, the number of neurites decreased, the length of cells decreased and the cell survival rate decreased. The levels of NCAM mRNA in 40 and 80μg / ml groups were significantly lower than those in control group, while the NCAM mRNA levels in 20μg / ml group were not significantly different from those in control group Certain dose-response relationship, and with the fluorine concentration increased and decreased. The protein level of NCAM-180 in 40 and 80μmg / ml groups was significantly lower than that in control group, the expression level of NCAM-140 protein in each group was significantly lower than that in control group, and the level of NCAM-120 protein in 80μg / ml group was significantly Lower than the control group. Conclusion Fluoride can inhibit the growth and survival of hippocampal cells and decrease the expression of NCAM mRNA and protein. Fluoride may be one of the neurotoxic sites in developing hippocampal damage.