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Early Evidence of Global DNA Methylation and Hydroxymethylation Changes in Rat Kidneys Consequent to Hyperoxaluria-Induced Renal Calcium Oxalate Stones

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Abstract

Calcium supersaturation in urine during hyperoxaluria can lead to renal calcium oxalate (CaOx) stone deposition; a condition termed as nephrolithiasis. Recent genetic studies indicate the possibility of epigenetic alterations during nephrolithiasis. We aimed to study the influence of renal CaOx stone formation on the global levels of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) in hyperoxaluric rat kidney. In this study, twelve male Wistar rats were divided equally into two groups (control and experimental). Animals in the experimental group received 2.5% (w/v) sodium oxalate in 5 mL of sterile water, every day, orally for four weeks to induce hyperoxaluria. At the end of four weeks, urine parameters, serum biochemistry and renal histopathological changes were evaluated. Global 5mC and 5hmC levels were analysed using enzyme-linked immune sorbent assay (ELISA), and renal mRNA expression of DNMTs and TET genes were also determined. Urine parameters and serum biochemistry showed that rats fed with 2.5% (w/v) sodium oxalate orally for four weeks developed hyperoxaluria (p < 0.05). Histopathological evaluation of hyperoxaluric rat kidneys showed the deposition of CaOx crystals and marked tubular injury. ELISA showed significantly high levels of 5hmC (p < 0.0001) in the kidneys of rats with renal CaOx stones, whereas 5mC was only slightly significant (p < 0.05) as compared to the control rats. mRNA expression of TET 2 a regulator of DNA demethylation process, was found to be significantly upregulated (p < 0.01) in the rats with CaOx kidney stones. This preliminary study showed some early evidence of epigenetic modifications being influenced by hyperoxaluria-induced renal CaOx stones, which may likely contribute to the transcriptional regulation during CaOx nephrolithiasis.

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ACKNOWLEDGMENTS

The authors would like to thank Anubhav Chakraborty from Vellore Institute of Technology for his assistance in language and grammatical correction. The work was funded by grants from the Indian Council of Medical Research (no. 5/9/1049/2013-Nut.).

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Authors and Affiliations

  1. Renal Research Lab., Centre for Bio-Medical Research, School of Bio-Sciences and Technology, Vellore Institute of Technology, 632014, Vellore, Tamil Nadu, India

    Yogita Mehra &  Pragasam Viswanathan

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  1. Yogita Mehra
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  2. Pragasam Viswanathan
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Correspondence to Pragasam Viswanathan.

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Conflict of interest. The authors declare that they have no conflicts of interest.

Statement on the welfare of animals. The protocols for animal experimental procedure used in this study were approved by the Institutional Animal Ethics Committee (Registration no. 1333/c/10/CPCSEA; Approval no. VIT/IAEC/15/ Sep2/25). All procedures performed in this study involving animals were carried out according to the guidelines set by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India.

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Yogita Mehra, Pragasam Viswanathan Early Evidence of Global DNA Methylation and Hydroxymethylation Changes in Rat Kidneys Consequent to Hyperoxaluria-Induced Renal Calcium Oxalate Stones. Cytol. Genet. 56, 458–465 (2022). https://doi.org/10.3103/S0095452722050085

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  • DOI: https://doi.org/10.3103/S0095452722050085

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