当前位置: X-MOL 学术Pathobiology › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Upregulation of Mitochondrial Transcription Factor A Promotes the Repairment of Renal Tubular Epithelial Cells in Sepsis by Inhibiting Reactive Oxygen Species-Mediated Toll-Like Receptor 4/p38MAPK Signaling
Pathobiology ( IF 5 ) Pub Date : 2019-01-01 , DOI: 10.1159/000501789
Xin-Gui Dai 1, 2 , Tao Li 2 , Wei-Bo Huang 3 , Zhen-Hua Zeng 4 , Qiong Li 2 , Yang Yang 2 , Ze-Peng Duan 2 , Yu-Jing Wang 2 , Yu-Hang Ai 5
Affiliation  

Background: Mitochondrial transcription factor A (TFAM) plays multiple pathophysiologic roles in mitochondrial DNA (mtDNA) maintenance. However, the role of TFAM in sepsis-induced acute kidney injury (AKI) remains largely unknown. Methods: Lipopolysaccharide (LPS) treatment of HK-2 cells mimics the in vitro model of AKI inflammation. pcDNA3.1 plasmid was used to construct pcDNA3.1-TFAM. sh-TFAM-543, sh-TFAM-717, sh-TFAM-765, sh-TFAM-904 and pcDNA3.1-TFAM were transfected into HK-2 cells using Lipofectamine 2000. MtDNA transcriptional levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay was performed to assess the cell viability. Changes in reactive oxygen species (ROS) and mitochondrial membrane potential in HK-2 cells were detected using the corresponding kits. Immunofluorescence experiment was used to investigate the displacement of TFAM. mRNA and protein expression levels of TFAM and its related genes were measured by qRT-PCR and western blot respectively. Mice in sepsis were administered cecal ligation and puncture surgery. Results: LPS treatment was a non-lethal influencing factor, leading to the upregulation of ROS levels and downregulation of mtDNA copy number and NADH dehydrogenase subunit-1 (ND1) expression, and caused damage to the mitochondria. As the LPS treatment time increased, TFAM was displaced from the periphery of the nucleus to cytoplasm. TFAM reduced ROS and P38MAPK levels by inhibiting toll-like receptor 4 (TLR4) expression, ultimately inhibiting inflammation and repairing mtDNA. Conclusions: Our results indicate that TFAM repairs mtDNA by blocking the TLR4/ROS/P38MAPK signaling pathway in inflammatory cells, thereby repairing septic tubular epithelial cells, and TFAM may serve as a new target for sepsis therapy.

中文翻译:

线粒体转录因子 A 的上调通过抑制活性氧介导的 Toll 样受体 4/p38MAPK 信号传导促进脓毒症肾小管上皮细胞的修复

背景:线粒体转录因子 A (TFAM) 在线粒体 DNA (mtDNA) 维持中发挥多种病理生理作用。然而,TFAM 在败血症引起的急性肾损伤 (AKI) 中的作用仍然未知。方法:脂多糖 (LPS) 处理 HK-2 细胞模拟 AKI 炎症的体外模型。pcDNA3.1质粒用于构建pcDNA3.1-TFAM。使用 Lipofectamine 2000 将 sh-TFAM-543、sh-TFAM-717、sh-TFAM-765、sh-TFAM-904 和 pcDNA3.1-TFAM 转染到 HK-2 细胞中。通过实时定量检测 MtDNA 转录水平聚合酶链反应 (qRT-PCR)。进行 3-(4,5)-二甲基噻唑 (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) 测定以评估细胞活力。使用相应的试剂盒检测 HK-2 细胞中活性氧 (ROS) 和线粒体膜电位的变化。免疫荧光实验用于研究TFAM的置换。通过qRT-PCR和蛋白质印迹分别测量TFAM及其相关基因的mRNA和蛋白质表达水平。对败血症小鼠进行盲肠结扎和穿刺手术。结果:LPS处理是非致死性影响因素,导致ROS水平上调、mtDNA拷贝数和NADH脱氢酶亚基1(ND1)表达下调,并导致线粒体损伤。随着 LPS 处理时间的增加,TFAM 从细胞核的外围转移到细胞质。TFAM 通过抑制 Toll 样受体 4 (TLR4) 表达降低 ROS 和 P38MAPK 水平,最终抑制炎症和修复 mtDNA。结论:我们的结果表明,TFAM 通过阻断炎症细胞中的 TLR4/ROS/P38MAPK 信号通路来修复 mtDNA,从而修复脓毒症肾小管上皮细胞,并且 TFAM 可以作为脓毒症治疗的新靶点。
更新日期:2019-01-01
down
wechat
bug