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

Autor:	Ze-Peng Duan, Xin-Gui Dai, Qiong Li, Yu-Jing Wang, Zhenhua Zeng, Yuhang Ai, Tao Li, Yang Yang, Wei-Bo Huang
Rok vydání:	2019
Předmět:	Inflammation Mitochondrion p38 Mitogen-Activated Protein Kinases Cell Line Pathology and Forensic Medicine Mitochondrial Proteins Mice 03 medical and health sciences 0302 clinical medicine Downregulation and upregulation Sepsis medicine Animals Humans Viability assay Molecular Biology Toll-like receptor Chemistry Epithelial Cells Cell Biology General Medicine Transfection TFAM Cell biology DNA-Binding Proteins Mice Inbred C57BL Toll-Like Receptor 4 Kidney Tubules 030220 oncology & carcinogenesis TLR4 medicine.symptom Reactive Oxygen Species Signal Transduction Transcription Factors 030215 immunology
Zdroj:	Pathobiology. 86:263-273
ISSN:	1423-0291 1015-2008
DOI:	10.1159/000501789
Popis:	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.
Databáze:	OpenAIRE
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