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International journal of molecular sciences2023; 24(19); 14515; doi: 10.3390/ijms241914515

Identification of Potential miRNA Biomarkers to Detect Hydrocortisone Administration in Horses.

Abstract: Circulating microRNAs (miRNAs) are stable in bodily fluids and are potential biomarkers of various diseases and physiological states. Although several studies have been conducted on humans to detect drug doping by miRNAs, research on drugs and miRNAs in horses is limited. In this study, circulating miRNAs in horses after hydrocortisone administration were profiled and variations in miRNAs affected by hydrocortisone administration during endogenous hydrocortisone elevation were examined. The miRNAs were extracted from thoroughbred horse plasma before and after hydrocortisone administration and subjected to small RNA sequencing and reverse transcription quantitative PCR (RT-qPCR). RT-qPCR validation was performed for the 20 miRNAs that were most affected by hydrocortisone administration. The effects of elevated endogenous hydrocortisone levels due to exercise and adrenocorticotropic hormone administration were also confirmed. The validation results showed that approximately half of the miRNAs showed the same significant differences as those obtained using small RNA sequencing. Among the twenty miRNAs, two novel miRNAs and miR-133a were found to vary differently between exogenous hydrocortisone administration and endogenous hydrocortisone elevation. This study provides basic knowledge regarding the circulating miRNA profile of horses after hydrocortisone administration and identifies three miRNAs that could potentially be used as biomarkers to detect hydrocortisone administration.
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Publication Date: 2023-09-25 PubMed ID: 37833961PubMed Central: PMC10572364DOI: 10.3390/ijms241914515Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates whether certain small RNA molecules known as miRNAs can be used as biomarkers to detect when horses have been administered hydrocortisone. The study found that three specific miRNAs showed significant changes after hydrocortisone administration, signifying that they could be used as potential biomarkers.

Objective of the Research

  • This study set out to identify biomarkers that could reliably detect the administration of hydrocortisone in horses. By profiling circulating miRNAs in horses after hydrocortisone use, the scientists hoped to provide basic knowledge for its identification and to set a foundation for future research.

Conducting the Research

  • For effective detection, the researchers had to extract miRNAs from thoroughbred horses’ plasma track before and after administering hydrocortisone. They then subjected these miRNAs to small RNA sequencing and reverse transcription quantitative PCR (RT-qPCR).

Validation Using RT-qPCR

  • The researchers validated the top 20 miRNAs which showed the most change after administering hydrocortisone through RT-qPCR technique. This process is crucial in solidifying the findings of the RNA sequencing step.

Effects of Elevated Endogenous Hydrocortisone Levels

  • The study also confirmed the effects of raised internal hydrocortisone levels as a result of exercise and adrenocorticotropic hormone administration to provide a broad context for their findings.

Results of the Study

  • The results showed significant differences in about half of the miRNAs examined, which was consistent with what they obtained from RNA sequencing.
  • The study also identified three miRNAs (two novel ones and miR-133a) that demonstrated a different variance between exogenous hydrocortisone administration (introduced from outside) and endogenous hydrocortisone elevation (caused internally).

Significance of the research

  • The findings from this research contribute fundamental knowledge about the miRNA profile in horses after the administration of hydrocortisone.
  • It identified three potential miRNA biomarkers that could be used in future to detect hydrocortisone administration in horses, a breakthrough in the field of veterinary pharmacology.

Cite This Article

APA
Kikuchi M, Ishige T, Minamijima Y, Hirota KI, Nagata SI, Tozaki T, Kakoi H, Ishiguro-Oonuma T, Kizaki K. (2023). Identification of Potential miRNA Biomarkers to Detect Hydrocortisone Administration in Horses. Int J Mol Sci, 24(19), 14515. https://doi.org/10.3390/ijms241914515

Publication

International journal of molecular sciences
ISSN: 1422-0067
NlmUniqueID: 101092791
Country: Switzerland
Language: English
Volume: 24
Issue: 19
PII: 14515

Researcher Affiliations

Kikuchi, Mio
  • Genetic Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsurutamachi, Utsunomiya 320-0851, Tochigi, Japan.
  • Graduate School of Veterinary Sciences, Iwate University, 3-18-8 Ueda, Morioka 020-8550, Iwate, Japan.
Ishige, Taichiro
  • Genetic Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsurutamachi, Utsunomiya 320-0851, Tochigi, Japan.
Minamijima, Yohei
  • Drug Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsurutamachi, Utsunomiya 320-0851, Tochigi, Japan.
Hirota, Kei-Ichi
  • Genetic Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsurutamachi, Utsunomiya 320-0851, Tochigi, Japan.
Nagata, Shun-Ichi
  • Genetic Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsurutamachi, Utsunomiya 320-0851, Tochigi, Japan.
Tozaki, Teruaki
  • Genetic Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsurutamachi, Utsunomiya 320-0851, Tochigi, Japan.
Kakoi, Hironaga
  • Genetic Analysis Department, Laboratory of Racing Chemistry, 1731-2 Tsurutamachi, Utsunomiya 320-0851, Tochigi, Japan.
Ishiguro-Oonuma, Toshina
  • Graduate School of Veterinary Sciences, Iwate University, 3-18-8 Ueda, Morioka 020-8550, Iwate, Japan.
Kizaki, Keiichiro
  • Graduate School of Veterinary Sciences, Iwate University, 3-18-8 Ueda, Morioka 020-8550, Iwate, Japan.

MeSH Terms

  • Humans
  • Horses / genetics
  • Animals
  • MicroRNAs / genetics
  • Hydrocortisone / pharmacology
  • Biomarkers
  • Circulating MicroRNA / genetics
  • Real-Time Polymerase Chain Reaction

Grant Funding

  • 2020-2022 / Japan Racing Association

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

This article has been cited 2 times.
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