核呼吸因子1在心肌细胞损伤中的保护作用  

The protect role of nuclear respiratory factor 1 in mitochondrial damage in cardiac muscles

作　　者：秦国华[1,2] 王瑞霞[1] 徐志芳[1,3] 桑楠[1] QIN Guo-hua;WANG Rui-xia;XU Zhi-fang;SANG Nan(College of Environmental Science and Resources, Shanxi University Taiyuan 030006, China;Guangzhou Key Laboratory of Environmental Exposure and Health, School of Environment,Jinan University, Guangzhou 510632, China;Shanxi Academy for Environmental Planning, Taiyuan 030001, China)

机构地区：[1]山西大学环境与资源学院,山西太原030006 [2]暨南大学环境学院,广州市环境暴露与健康重点实验室,广东广州510632 [3]山西省环境规划院,山西太原030001

出　　处：《中国环境科学》2017年第4期1556-1561,共6页China Environmental Science

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)

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細胞,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細胞中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.甶此说明: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的表达进而増加細胞内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,掲示NRF1在调节NaHSO_3诱导的线粒体损伤过程中有重要意义.We constructed a NRF1-pcDNA3.1plasmid and transfected it in H9C2cells to explore the effects of NRF1in sodium bisulfite-induced mitochondrial damage in cardiac muscles of rats.H9C2cells were treated with100μmol/LNaHSO3for24h after NRF1-pcDNA3.1or pcDNA3.1transfection.Protein levels of NRF1and TFAM were measured by Western blotting.The amount of ATP was detected by the chemiluminescence method.It showed that NaHSO3treatment for24h resulted in significant decreases of NRF1and TFAM protein expression and ATP amount.The protein levels of NRF1and TFAM were significantly elevated after NRF1-pcDNA3.1plasmid transfection,companied by a significant increase of ATP amounts.NRF1and TFAM protein expressions and ATP amount significantly increased after NaHSO3exposure for24h in NRF1over-expression group,compared with the pcDNA3.1-transfection group.It implied that over-expression of NRF1might regulate TFAM to increase intracellular ATP production.In addition,over-expression of NRF1alleviated the dysfunction of mitochondrial ATP caused by NaHSO3exposure.NRF1plays an important role in the regulation of NaHSO3-induced mitochondrial damage.

关 键 词：NRF1 TFAM 亚硫酸氢钠 质粒构建 

分 类 号：X503[环境科学与工程—环境工程]