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[目的]探讨谷氨酸NMDA受体拮抗剂D-2-氨基-5-磷酰基戊酸(D-AP5)对氟砷联合染毒后原代海马神经元突触相关蛋白表达的影响。[方法]将体外培养的海马神经元分为4组,空白对照组、氟砷联合染毒组(0.4μg/m L氟化钠+0.1μg/m L亚砷酸钠)、D-AP5组(25μmol/L D-AP5)和D-AP5+氟砷联合染毒组(0.4μg/m L氟化钠+0.1μg/m L亚砷酸钠+25μmol/L D-AP5)。各组海马神经元置于37℃、含有体积分数5%CO_2培养箱中培养24 h。把培养好的各组海马神经元用CCK8检测活性,用蛋白免疫印迹法检测海马神经元中NR2B、SYN及GAP43突触相关蛋白的表达水平。[结果]与空白对照组[(100.00±0.00)%]比较,氟砷联合染毒组[(75.81±6.27)%]、D-AP5+氟砷联合染毒组[(81.58±5.20)%]的细胞存活率均明显降低(P<0.05);与氟砷联合染毒组[(75.81±6.27)%]比较,D-AP5组[(98.78±3.53)%]、D-AP5+氟砷联合染毒组[(81.58±5.20)%]海马神经元存活率均明显升高(P<0.05)。氟砷联合染毒组(0.425±0.06)、D-AP5组(0.469±0.03)、D-AP5+氟砷联合染毒组(0.354±0.04)的NR2B相对表达量较空白对照组(0.572±0.05)均有所下降(P<0.05),D-AP5+氟砷联合染毒组(0.354±0.04)受体蛋白NR2B表达较氟砷联合染毒组(0.425±0.06)降低(P<0.05);氟砷联合染毒组与D-AP5+氟砷联合染毒组的SYN和GAP43相对表达量均较空白对照组低(P<0.05)。而D-AP5组与D-AP5+氟砷联合染毒组的SYN和GAP43表达较氟砷联合染毒组均升高(P<0.05)。[结论]氟砷联合染毒能够下调突触相关蛋白SYN及GAP-43表达水平,谷氨酸NMDA受体拮抗剂D-AP5可以抑制氟砷联合染毒对海马神经元突触相关蛋白表达的影响。
[Objective] To investigate the effect of D-2-amino-5-phosphoryl-pentanoic acid (D-AP5), a glutamate NMDA receptor antagonist, on the expression of synaptic proteins in primary cultured hippocampal neurons after exposure to fluvastatin. [Methods] The hippocampal neurons cultured in vitro were divided into four groups: blank control group, combination of fluoride-arsenic exposure group (0.4μg / mL sodium fluoride + 0.1μg / mL sodium arsenite), D-AP5 group (25μmol / L D-AP5) and D-AP5 + fluoride-arsenic combined treatment group (0.4μg / m L sodium fluoride + 0.1μg / m L sodium arsenite + 25μmol / L D-AP5). Hippocampal neurons in each group were incubated at 37 ℃ for 24 h with a volume fraction of 5% CO 2. The cultured hippocampal neurons were detected by CCK8 and the expression of NR2B, SYN and GAP43 synapse-related proteins in hippocampal neurons were detected by Western blotting. [Results] Compared with the blank control group [(100.00 ± 0.00)%], there was no significant difference between the fluoride-arsenic combined treatment group [(75.81 ± 6.27)%] and the D-AP5 + (98.78 ± 3.53)%], D-AP5 + fluoride-arsenite combined with fluoride-arsenic exposure group [(75.81 ± 6.27)%] Group [(81.58 ± 5.20)%] hippocampal neurons survival rates were significantly increased (P <0.05). Compared with blank control group (0.572 ± 0.05), the relative expression of NR2B in fluoride-arsenic combined treatment group (0.425 ± 0.06), D-AP5 group (0.469 ± 0.03) and D-AP5 + fluoride-arsenic combined treatment group (0.354 ± 0.04) (P <0.05). The expression of NR2B receptor of 0.354 ± 0.04 receptor protein in D-AP5 + fluoride-arsenite combined treatment group was significantly lower than that in control group (0.425 ± 0.06) (P <0.05) The relative expression of SYN and GAP43 in combination group and D-AP5 + fluoride-arsenic combination group were lower than those in blank control group (P <0.05). However, SYN and GAP43 expression in D-AP5 group and D-AP5 + fluoride-arsenic combination group were higher than that in fluoride-arsenic combination group (P <0.05). [Conclusion] The combination of fluoride and arsenic can down-regulate the expression of SYN and GAP-43, and the glutamate NMDA receptor antagonist D-AP5 can inhibit the expression of synaptotagmin in hippocampus influences.