Tian Z Y, Chen J W, Zhou G Y, Li P, Zhao Q, Luo C, Zhang S, Wang A G
Department of Occupational and Environmental Health, School of Public Health, MOE Key Lab of Environment and Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2018 Oct 20;36(10):721-727. doi: 10.3760/cma.j.issn.1001-9391.2018.10.001.
To explore the role of mitochondrial biogenesis and the neuroprotective mechanism of resveratrol in fluoride neurotoxicity. SH-SY5Y cells in exponential phase were treated with different concentrations (20, 40, 60 mg/L) of sodium fluoride (NaF) for 24 h. Co-treatment with 60 mg/L NaF, 20 μmol/L resveratrol (RSV) was administrated in the resveratrol intervene trial. Western blotting was used to determine the expression levels of mitochondrial biogenesis key regulating factor of peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) , nuclear respiratory factor 1 (NRF1) and mitochondrial transcription factor A (TFAM) in SH-SY5Y cells. The mRNA levels of PGC-1α, NRF1 and TFAM were determined by Quantitative Real-time PCR in SH-SY5Y cells, as well as the relative mitochondrial DNA (mtDNA) contents and mRNA expression of mitochondrial respiratory chain complexes subunit CO1 and ATP8. Flow cytometry was used to determine mitochondrial membrane potential in SH-SY5Y cells. Both the protein and mRNA levels of PGC-1α, NRF1 and TFAM were decresed after 60 mg/L NaF treatment in SH-SY5Y cells (<0.05) . The relative mtDNA contents and mRNA expression of complexes subunit CO1 and ATP8 were also significantly decreased compared with control (<0.05) . Mitochondrial membrane potential were also significantly decreased after 60 mg/L NaF treatment in SH-SY5Y cells (<0.05) . Compared with 60 mg/L NaF group, the protein and mRNA levels of PGC-1α, NRF1 and TFAM in 20 μmol/L RSV+60 mg/L NaF group were significantly increased (<0.05) . The relative mtDNA contents, mitochondrial membrane potential and mRNA levels of complexes subunit CO1 and ATP8 in 20 μmol/L RSV+60 mg/L NaF group were also significantly higher than that in 60 mg/L NaF group (<0.05) . Resveratrol may alleviate the fluoride-induced mitochondrial biogenesis dysfunction in SH-SY5Y cells.
探讨线粒体生物合成的作用以及白藜芦醇在氟神经毒性中的神经保护机制。将指数生长期的SH-SY5Y细胞用不同浓度(20、40、60mg/L)的氟化钠(NaF)处理24小时。在白藜芦醇干预试验中,用60mg/L NaF与20μmol/L白藜芦醇(RSV)共同处理。采用蛋白质免疫印迹法检测SH-SY5Y细胞中过氧化物酶体增殖物激活受体γ共激活因子1α(PGC-1α)、核呼吸因子1(NRF1)和线粒体转录因子A(TFAM)等线粒体生物合成关键调节因子蛋白表达水平。采用实时荧光定量PCR法检测SH-SY5Y细胞中PGC-1α、NRF1和TFAM的mRNA水平,以及线粒体DNA(mtDNA)相对含量和线粒体呼吸链复合物亚基CO1和ATP8的mRNA表达。采用流式细胞术检测SH-SY5Y细胞线粒体膜电位。60mg/L NaF处理后,SH-SY5Y细胞中PGC-1α、NRF1和TFAM的蛋白和mRNA水平均降低(<0.05)。与对照组相比,mtDNA相对含量以及复合物亚基CO1和ATP8的mRNA表达也显著降低(<0.05)。60mg/L NaF处理后,SH-SY5Y细胞线粒体膜电位也显著降低(<0.05)。与60mg/L NaF组相比,20μmol/L RSV+60mg/L NaF组中PGC-1α、NRF1和TFAM的蛋白和mRNA水平显著升高(<0.05)。20μmol/L RSV+60mg/L NaF组的mtDNA相对含量、线粒体膜电位以及复合物亚基CO1和ATP8的mRNA水平也显著高于60mg/L NaF组(<0.05)。白藜芦醇可能减轻氟诱导的SH-SY5Y细胞线粒体生物合成功能障碍。