摘要
目的通过研究神经细胞线粒体改变探索氟中毒致神经损伤的潜在生物学机制。方法将SH-SY5Y细胞设置为对照组和20、40、60mg/LNaF染毒剂量组,检测线粒体跨膜电位(ΔΨm)及线粒体内活性氧(MitoROS)水平,通过Westernblot法检测线粒体动力学和自噬相关蛋白(Miro1、ATG5和PINK1)及凋亡相关蛋白(PARP、cleaved Caspase-3和Bcl-2)的表达情况,细胞免疫荧光法检测Miro1、ATG5和PINK1蛋白与线粒体共定位情况,流式细胞术检测细胞凋亡。结果随着NaF浓度增加,SH-SY5Y细胞ΔΨm降低(P<0.01),MitoROS水平升高(P<0.05);线粒体转运蛋白Miro1表达升高(P<0.05),自噬相关蛋白ATG5(P<0.05)和PINK1(P<0.05)表达均升高;细胞早期凋亡率和总凋亡率均升高(均P<0.05),凋亡相关蛋白PARP(P<0.05)和cleavedCaspase-3(P<0.05)表达升高,抗凋亡蛋白Bcl-2表达降低(P<0.05)。结论高剂量氟可促进线粒体的氧化应激、转运和自噬,并诱导神经元凋亡。
Objective To explore the potential mechanism of the effect of fluoride on nerve injury by studying the changes of mitochondria in neural cells.Methods SH-SY5 Y cells were treated with NaF in vitro(0 mg/L,20 mg/L,40 mg/L and 60 mg/L).Western blotting,immunofluorescence,and cell flow cytometry methods were utilized to detect the expression levels of mitochondrial dynamics-related and autophagy-related proteins(Miro1,ATG5 and PINK1),and apoptosis-related proteins(PARP,cleaved Caspase-3 and Bcl-2),the colocalization of mitochondrial dynamics-related proteins and mitochondria,mitochondrial membrane potential(ΔΨm)and oxidation level(MitoROS),and apoptosis rate,respectively.Results With increasing dose of fluoride exposure,ΔΨm reduced(P<0.01)and MitoROS content increased(P<0.05).In addition,with increasing dose of fluoride exposure,the expression of mitochondrial transporter Miro1(P<0.05),autophagy-related proteins ATG5(P<0.05)and PINK1(P<0.05)were upregulated.What’s more,with increasing dose of fluoride exposure,both the early and total apoptosis rates were increased(both P<0.05),and the expression of apoptosis-related proteins PARP and cleaved Caspase-3 upregulated(both P<0.05),while the expression of Bcl-2 was significantly downregulated(P<0.05).Conclusion Fluoride could induce neurotoxicity by interfering mitochondrial oxidative stress,transfer and autophagy.
作者
李新颖
于星辰
张舜
王爱国
刘莉
Li Xinying;Yu Xingchen;Zhang Shun(Department of Epidemiology and Biostatistics,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China;MOE Key Lab of Environment and Health,Department of Occupational and Environmental Health,School of Public Health,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2021年第5期555-560,共6页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81430076)。
关键词
氟
线粒体
神经损伤
fluoride
mitochondria
nerve injury
