摘要
目的探索亮氨酸羧基甲基转移酶1(LCMT1)高表达对锰诱导神经细胞损伤的拮抗作用。方法利用脂质体将LCMT1-PCDNA 3.0和PCDNA 3.0质粒分别转染小鼠神经母细胞瘤细胞(N2a细胞),通过G418筛选稳定高表达LCMT1的LCMT1-N2a细胞和PCDNA-N2a细胞,Western blot检测显示LCMT1蛋白过表达情况。将LCMT1-N2a细胞和PCDNA-N2a细胞分别暴露于空白培养基(对照组)和不同浓度氯化锰(250、500、1000、2000μmol/L)12 h后以刃天青法检测细胞活性。随后,用流式细胞仪检测染锰(0、500、1000μmol/L)12 h后细胞的凋亡率。结果LCMT1蛋白在LCMT1-PCDNA 3.0细胞中的表达高于对照组且差异具有统计学意义(P<0.05);转染后细胞的形态与转染前无明显变化。锰暴露可使PCDNA-N2a细胞活性下降且有剂量-反应关系,相同浓度锰暴露下LCMT1-N2a细胞存活率高于PCDNA3.0 N2a(P<0.05)。PCDNA3.0 N2a细胞的凋亡率随着染锰浓度增加而升高且存在剂量-反应关系;在相同的染锰剂量下,LCMT1-PCDNA3.0 N2a细胞的凋亡率低于PCDNA3.0 N2a细胞(P<0.05)。结论LCMT1高表达可以拮抗锰致N2a细胞细胞活性的下降和细胞凋亡。
Objective To explore the antagonism of overexpression of leucine carboxyl methyltransferase 1(LCMT1)to nerve cell damage induced by manganese.Methods LCMT1-N2 a and PCDNA-N2 a cells transfected with LCMT1 plasmid and control plasmid was verified by Western blot assays,and screened by G418,then were exposed to 0,250,500,1000,2000μmol/L manganese chloride.After 12 h-treatment,cell viability was detected by alamar Blue Assay.The apoptosis of cells treated with0,500,1000μmol/L manganese chloride for 12 h was determined by flow cytometry.Results LCMT1 protein in LCMT1 overexpressed cells was higher than control cells,and the difference was statistically significant.There was no significant change in cell morphology after transfection.Manganese exposure decreased the activity of PCDNA-N2 a cells in a dose-response relationship.With the same manganese exposure condition,the survival rates of LCMT1-N2 a cells were significantly higher than PCDNA-N2 a cells(P<0.05).The apoptosis rate of PCDNA-N2 a cells increased with the increase of manganese concentration in a dose-dependent relationship.With the same exposure conditions,the apoptosis rate of LCMT1-N2 a cells was significantly lower than PCDNA-N2 a cells(P<0.05).Conclusion Overexpression of LCMT1 can enhance N2 a cells resistance to manganese-induced cell activity damage and apoptosis.
作者
韦兰成
徐怡璐
石倩倩
沈颖慧
黄丽媛
陆建勇
李习艺
唐深
王新航
陆彩玲
WEI Lan-cheng;XU Yi-lu;SHI Qian-qian;SHEN Ying-hui;HUANG Li-yuan;LU Jian-yong;LI Xi-yi;TANG Shen;WANG Xin-hang;LU Cai-ling(School of Public Health,Guangxi Medical University,Nanning,Guangxi 530021,China;不详)
出处
《环境与健康杂志》
CAS
北大核心
2021年第3期197-200,283,共5页
Journal of Environment and Health
基金
国家自然科学基金(81760576,21567006)
