Identification of stable housekeeping genes in mouse liver for studying carbon tetrachloride-induced injury and cellular senescence.

Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE

Original Publication: London : Nature Publishing Group, copyright 2011-

Cellular Senescence*/genetics ; Carbon Tetrachloride*/toxicity ; Genes, Essential* ; Liver*/metabolism ; Liver*/pathology ; Liver*/drug effects; Animals ; Mice ; Chemical and Drug Induced Liver Injury/genetics ; Chemical and Drug Induced Liver Injury/metabolism ; Chemical and Drug Induced Liver Injury/pathology ; Male ; Gene Expression Profiling/methods ; Disease Models, Animal ; Mice, Inbred C57BL

Acute liver injury (ALI) presents a challenging problem worldwide, prompting extensive research efforts. Cellular senescence has been found to be induced following ALI, and targeting cellular senescence has shown therapeutic potential. Therefore, understanding the expression of senescence-related genes in ALI can help to explore pathogenesis and treatment of this common disease. Carbon tetrachloride (CCl 4 ) is commonly used to study ALI. Although polymerase chain reaction (PCR) is a convenient and economical molecular biology technique widely used in basic medicine, research on selecting suitable reference genes to obtain objective and reproducible PCR data is scarce. Moreover, evidence supporting the choice of reference genes for experimental studies of CCl4-induced ALI and hepatic senescence in mice is limited. In this study, we obtained murine livers at four time points (0, 12, 24, and 48 h) following CCl4 treatment. We used five algorithms (geNorm, BestKeeper, NormFinder, delta Ct, and RefFinder) to rank 12 candidate genes in real-time reverse-transcription quantitative PCR (RT-qPCR) experiments. Focusing on cellular senescence in this model, we adopted four senescence-associated secretory phenotype (SASP) genes (Il6, Il1b, Ccl2, and Ccl5) as target genes. Our results confirmed Gapdh and Tbp as suitable reference genes in murine CCl 4 -induced ALI models. Furthermore, we provide a table of published studies recommending reference genes for various liver disease models. This study provides a valuable reference for enhancing the reliability and reproducibility of ALI molecular findings.
(© 2024. The Author(s).)

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2020YFE0204300 National Key Research and Development Program of China; 2020YFE0204300 National Key Research and Development Program of China; 2020YFE0204300 National Key Research and Development Program of China; 2022ZFJH003 Fundamental Research Funds for the Central Universities; 2022ZFJH003 Fundamental Research Funds for the Central Universities; 2022ZFJH003 Fundamental Research Funds for the Central Universities

Keywords: Carbon Tetrachloride; Cellular senescence; Mouse; Polymerase chain reaction; Reference gene

CL2T97X0V0 (Carbon Tetrachloride)

Date Created: 20241103 Date Completed: 20241103 Latest Revision: 20241106

20250114

PMC11532458

10.1038/s41598-024-78183-y

39489763