核呼吸因子1在心肌细胞损伤中的保护作用

发布时间：2018-04-14 22:18

本文选题：NRF + TFAM　；参考：《中国环境科学》2017年04期

【摘要】：构建含有核呼吸因子1(NRF1)cDNA全长的质粒NRF1-pcDNA3.1,用该质粒转染H9C2}0胞,探讨NRF1在NaHSO_3诱导心肌细胞线粒体损伤中的作用.用空质粒或NRF1-pcDNA3.1转染H9C2细胞,100pmol/L的NaHSO_3处理转染后的H9C2心肌细胞24h后,首先采用Western blot免疫印迹法检测H9C2心肌细胞中NRF1和线粒体转录因子A(TFAM)蛋白的表达情况.其次,采用化学发光法检测}0胞中ATP含量,考察NRF1对NaHSO_3诱导H9C2心肌细胞线粒体损伤的影响.结果表明,与未染毒组相比,空质粒转染组NaHSO_3染毒24h后,NRF1和TFAM蛋白的表达及ATP含量均降低;而NRF1-pcDNA3.1转染显著上调了NRF1和TFAM蛋白表达和细胞内ATP含量;NRF1-pcDNA3.1转染的H9C2细胞经过NaHSO_3染毒24h后,与空质粒转染后NaHSO_3染毒组相比,NRF1和TFAM的蛋白表达及ATP含量均极显著上升.u6此说明:NaHSO_3可降低H9C2细胞中NRF1和TFAM表达及ATP含量.NRF1的高表达可通过调控下游基因TFAM的表达进而X椉觹0胞内ATP的产生,并且NRF1的高表达可使暴露于NaHSO_3所引起的线粒体功能异常得到缓解,c彩綨RF1在调节NaHSO_3诱导的线粒体损伤过程中有重要意义.
[Abstract]:A full-length plasmid NRF1-pcDNA3.1 containing nuclear respiratory factor (1(NRF1)cDNA) was constructed and transfected into H9C2} 0 cell to investigate the role of NRF1 in mitochondrial damage induced by NaHSO_3.After treated with 100 pmol / L NaHSO_3 of H9C2 cells transfected with empty plasmids or NRF1-pcDNA3.1 for 24 hours, the expression of NRF1 and mitochondrial transcription factor (AtoTFAM) protein in H9C2 cardiomyocytes was detected by Western blot Western blotting.Secondly, the content of ATP in} 0 cells was detected by chemiluminescence assay to investigate the effect of NRF1 on mitochondrial damage induced by NaHSO_3 in H9C2 cardiomyocytes.The results showed that the expression of NRF1 and TFAM protein and the content of ATP decreased in the empty plasmid transfection group 24 hours after exposure to NaHSO_3 compared with the control group.However, NRF1-pcDNA3.1 transfection significantly up-regulated the expression of NRF1 and TFAM protein and the content of ATP in the cells. The H9C2 cells transfected with NRF1-pcDNA3.1 were exposed to NaHSO_3 for 24 hours.The protein expression and ATP content of NRF1 and TFAM increased significantly after transfection of NaHSO_3 with empty plasmid. U6. This indicates that: NaHSO3 can reduce the expression of NRF1 and TFAM and the high expression of ATP. NRF1 in H9C2 cells by regulating the expression of downstream gene TFAM.And then the production of X * 0 intracellular ATP,Moreover, the high expression of NRF1 can alleviate the mitochondrial dysfunction induced by NaHSO_3 exposure, and RF1 may play an important role in regulating the mitochondrial damage induced by NaHSO_3.
【作者单位】：山西大学环境与资源学院;暨南大学环境学院广州市环境暴露与健康重点实验室;山西省环境规划院;
【基金】：国家自然科学基金资助项目(21007036) 广州市环境暴露与健康重点实验室开放基金资助项目(GZKLEEH201612) 环境化学与生态毒理学国家重点实验室开放基金资助项目(KF2016-17)
【分类号】：R114

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