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Published on: October 7, 2016
Combination of mRNA of Repair-related Genes in Rat Skeletal Muscles for Wound Age Estimation
1School of Forensic Medicine, Shanxi Medical University, Taiyuan 030001, China.
Abstract:
Objective To investigate the estimation of early and mid-term wound age by a combination of four mRNAs, the DNA polymerase delta-interacting protein 3 （POLDIP3） mRNA, regulator of chromosome condensation 1 like （RCC1L） mRNA, proline-rich 5 （PRR5） mRNA, and ribonucleic acid export 1 （RAE1） mRNA in rats skeletal muscles. Methods The model of rat skeletal muscle contusion was established, and then contusion area muscle tissue was extracted 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44 and 48 h after injury. Histomorphological changes during the repair process after rat skeletal muscle contusion were observed. The relative expressions of Poldip3, Rcc1l, Prr5 and Rae1 mRNAs were detected by reverse transcription real-time quantitative polymerase chain reaction （RT-qPCR）. Different stages of wound age were classified by using the expression patterns of four genes at various time points after injury. The accuracy of the results was verified by Fisher discriminant analysis. Results Histomorphological results showed that the repair process after skeletal muscle contusion occurred with the prolonging of time. Through combination of the expression trends of the four kinds of mRNAs, the 48 h after injury could be divided into three periods, 4-12 h, 16-28 h and 32-48 h. The Fisher discrimination method showed that the classification accuracy rates of the three periods were 83.3%, 75.0% and 73.3%, respectively. Conclusion The classification discrimination based on the relative expression of every gene has a higher accuracy, and the accuracy of wound age estimation with combination of mRNA relative expressions is higher than that with a single indicator. By combining with Fisher discrimination method, this method can be used for early and mid-term wound age estimation.
Insights
This study estimates wound age in rat skeletal muscle using four specific mRNAs. Combining these mRNA expressions with Fisher discriminant analysis improves accuracy for early and mid-term wound age determination.
Area of Science:
- Forensic Science
- Molecular Biology
- Tissue Repair Mechanisms
Background:
- Accurate estimation of wound age is crucial in forensic investigations.
- Skeletal muscle contusions present unique challenges for post-injury aging.
- Gene expression patterns offer potential biomarkers for temporal analysis of tissue damage.
Purpose of the Study:
- To evaluate the efficacy of a combination of four mRNAs (POLDIP3, RCC1L, PRR5, RAE1) for estimating early and mid-term wound age in rat skeletal muscle.
- To correlate mRNA expression levels with histomorphological changes during skeletal muscle contusion repair.
- To validate the accuracy of this multi-gene mRNA approach using Fisher discriminant analysis.
Main Methods:
- Established a rat skeletal muscle contusion model.
- Collected muscle tissue at multiple time points (4-48 hours post-injury).
- Quantified relative mRNA expression of POLDIP3, RCC1L, PRR5, and RAE1 using RT-qPCR.
- Analyzed expression patterns and applied Fisher discriminant analysis for classification.
Main Results:
- Histomorphological analysis confirmed progressive repair over time.
- Combined mRNA expression trends allowed division of the 48-hour period into three distinct phases: 4-12h, 16-28h, and 32-48h.
- Fisher discriminant analysis yielded classification accuracies of 83.3% (4-12h), 75.0% (16-28h), and 73.3% (32-48h).
Conclusions:
- Relative gene expression analysis, particularly using a combination of POLDIP3, RCC1L, PRR5, and RAE1 mRNAs, provides a more accurate method for wound age estimation than single markers.
- The integration of this multi-mRNA approach with Fisher discriminant analysis demonstrates significant potential for reliable early and mid-term wound age determination in skeletal muscle injuries.
- This molecular approach offers a promising tool for forensic pathology and injury analysis.
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