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目的研究锰暴露不同时长对神经元细胞突触囊泡及其相关蛋白的影响。方法将体外培养的神经元,分别用100μM锰处理0、6、12、18、24 h后,观察细胞活力及培养液中LDH的释放量,测定SNARE复合物相关蛋白的基因及蛋白表达,以及活动性突触囊泡的释放。结果神经元细胞用100μM锰暴露不同时长后,神经元损伤逐渐加重(P<0.01);与对照组比较,Syntaxin 1A的基因及蛋白表达均未见明显改变(P>0.05),SNAP 25的基因和蛋白表达均逐渐下降(P<0.05),VAMP 2的基因和蛋白表达均逐渐升高(P<0.01),进而导致SNARE复合物蛋白形成先升高后下降的趋势,同时活动性突触囊泡的释放也出现相应改变。结论锰暴露可以时间依赖性的干扰SNARE复合物相关蛋白表达,减少了神经元细胞内SNARE复合物的形成,进而导致活动性突触囊泡减少,造成神经递质释放紊乱。
Objective To study the effects of different exposure durations on synaptic vesicles and their related proteins in neurons. Methods The neurons cultured in vitro were treated with 100μM manganese for 0, 6, 12, 18 and 24 hours respectively. The viability of cells and the release of LDH in culture medium were measured. The gene and protein expression of SNARE complex-related proteins were determined. Active synaptic vesicles release. Results Compared with the control group, the expression of Syntaxin 1A gene and protein had no significant change (P> 0.05), while those of SNAP 25 gene (P <0.05). The gene and protein expression of VAMP 2 increased gradually (P <0.01), which led to the first increase and then decrease of SNARE complex protein formation. Meanwhile, the activity of synaptic vesicles Bubble release also appeared to change accordingly. Conclusions Manganese exposure can interfere with the expression of SNARE complex-related proteins in a time-dependent manner and reduce the formation of SNARE complexes in neurons, resulting in the reduction of active synaptic vesicles and the disorder of neurotransmitter release.