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Updated: May 22, 2026
Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration
Published on: June 7, 2014
Reduction of DNA hydroxymethylation in the mouse kidney insulted by ischemia reperfusion
Ning Huang1, Li Tan, Zhanggang Xue
1Department of Anesthesiology, Zhongshan Hospital, Shanghai Medical College of Fudan University, 180 Fenglin Road, Shanghai 200032, PR China.
Abstract:
Ischemia reperfusion (IR) is a frequent pathological injury to the perioperative patients. The molecular mechanism underlying IR injury is still not well characterized. In this study, we investigated the effect of IR injury on DNA hydroxymethylation in mouse kidney. Dot blot and immunochemistry analysis showed that the global level of 5-hydroxymethylcytosine (5hmC) was reduced in mouse kidney insulted by IR; however, the 5-methylcytosine (5mC) level had no significant change. hMeDIP-qPCR validated that IR injury also decreased the 5hmC enrichment at promoter regions of Cxcl10 and Ifngr2 genes. RT-qPCR analysis revealed that the mRNA levels of Cxcl10 and Ifngr2 increased in IR-injured kidney. In addition, mRNA expression of both Tet1 and Tet2 but not Tet3 was dramatically downregulated in IR-injured kidney. Taken together, our data provided the first evidence that IR injury influences DNA hydroxymethylation and Tet gene expression in mouse kidney, which may contribute to the regulation of gene transcription during renal IR injury.
Insights
Ischemia reperfusion injury reduces DNA hydroxymethylation (5hmC) in mouse kidneys, impacting gene expression. This study reveals altered Tet gene activity, offering insights into renal injury mechanisms.
Area of Science:
- Molecular Biology
- Renal Pathophysiology
- Epigenetics
Background:
- Ischemia reperfusion (IR) injury is a common complication in perioperative patients.
- The precise molecular mechanisms driving IR injury remain incompletely understood.
- Epigenetic modifications, such as DNA hydroxymethylation, are increasingly recognized for their role in cellular responses to injury.
Purpose of the Study:
- To investigate the impact of IR injury on DNA hydroxymethylation patterns in the mouse kidney.
- To explore the relationship between IR-induced changes in DNA hydroxymethylation and gene expression.
- To examine the expression of Ten-eleven translocation (Tet) genes, which are involved in DNA demethylation, following IR injury.
Main Methods:
- Dot blot and immunochemistry to assess global 5-hydroxymethylcytosine (5hmC) and 5-methylcytosine (5mC) levels.
- Hydroxymethylated DNA immunoprecipitation quantitative PCR (hMeDIP-qPCR) to evaluate 5hmC enrichment at specific gene promoters.
- Reverse transcription quantitative PCR (RT-qPCR) to measure mRNA levels of target genes (Cxcl10, Ifngr2) and Tet family genes (Tet1, Tet2, Tet3).
Main Results:
- Global 5hmC levels significantly decreased in IR-injured mouse kidneys, while 5mC levels remained unchanged.
- IR injury led to reduced 5hmC enrichment at the promoter regions of Cxcl10 and Ifngr2.
- mRNA levels of Cxcl10 and Ifngr2 were upregulated, alongside a significant downregulation of Tet1 and Tet2 gene expression in IR-injured kidneys.
Conclusions:
- IR injury profoundly influences DNA hydroxymethylation in the kidney.
- Altered DNA hydroxymethylation and suppressed Tet gene expression (Tet1, Tet2) are associated with changes in gene transcription during renal IR injury.
- These findings provide novel insights into the epigenetic regulation of gene expression in the context of kidney IR injury.
