Estimation of Postmortem Interval using microRNA in the liver in aluminum phosphide deaths in rat model
Hussein, D., Hassan, D., Abdelhady, E., Hablas, M., Ali, M. (2025). Estimation of Postmortem Interval using microRNA in the liver in aluminum phosphide deaths in rat model. EKB Journal Management System, 6(4), 74-95. doi: 10.21608/ejmr.2025.386764.1835
Dina A. Hussein; Dalia Abd ELwahab Hassan; Ebthal Gamal Abdelhady; Mohamed Ghazy Attia Hablas; Motee R. Ali. "Estimation of Postmortem Interval using microRNA in the liver in aluminum phosphide deaths in rat model". EKB Journal Management System, 6, 4, 2025, 74-95. doi: 10.21608/ejmr.2025.386764.1835
Hussein, D., Hassan, D., Abdelhady, E., Hablas, M., Ali, M. (2025). 'Estimation of Postmortem Interval using microRNA in the liver in aluminum phosphide deaths in rat model', EKB Journal Management System, 6(4), pp. 74-95. doi: 10.21608/ejmr.2025.386764.1835
Hussein, D., Hassan, D., Abdelhady, E., Hablas, M., Ali, M. Estimation of Postmortem Interval using microRNA in the liver in aluminum phosphide deaths in rat model. EKB Journal Management System, 2025; 6(4): 74-95. doi: 10.21608/ejmr.2025.386764.1835

Estimation of Postmortem Interval using microRNA in the liver in aluminum phosphide deaths in rat model

Egyptian Journal of Medical Research
Volume 6, Issue 4, October 2025, Pages 74-95    PDF (1.06 M)
Document Type: Original Article
DOI: 10.21608/ejmr.2025.386764.1835
Authors
Dina A. Hussein* 1; Dalia Abd ELwahab Hassan1; Ebthal Gamal Abdelhady2; Mohamed Ghazy Attia Hablas3; Motee R. Ali1
1Forensic medicine and clinical toxicology department, Faculty of medicine, Beni-Suef University, Beni-Suef, Egypt
2Medical Biochemistry and Molecular Biology, Faculty of medicine, Beni-Suef University, Beni-Suef, Egypt
3Histology and Cell Biology, Faculty of Medicine, Suez University P.O. Box: 43221, Suez, Egypt
Abstract
Background: One of the main challenges in forensic pathology is determining the postmortem interval (PMI). The autolysis mechanism is exploited in recent PMI enhancements. Objective: To determine if the assessment of PMI can be achieved by utilizing the expression of the liver's microtubule-associated protein light chain3 (LC3) and Serine/threonine-protein kinase (mTOR) levels in liver specimens from aluminum phosphide toxicity. Methods: This was a cross-sectional study that included 70 rats, which were divided equally into 7 toxicity groups. The toxicity groups were administered aluminum phosphide suspension via oral lavage, after which the rats died within six hours. Immediately after death, liver tissues were collected from the groups at (0, 1, 2, 3, 4, 5, and 6 days postmortem). MDA (Malondialdehyde), CAT (catalase), and mTOR levels were measured in liver tissues using ELISA. The LC3 level was monitored using the GoTaq® 1-Step RT-qPCR. Organ samples were used for histopathological examination. Results: MDA and mTOR were upregulated with a statistically significant difference between the groups, while CAT and LC3 showed a statistically significant decrease between groups (P value <0.05). There was positive correlation between PMI and MDA and mTOR (P-value <0.001), while there was a negative correlation between PMI with CAT and LC3 (P-value <0.001). The linear regression analysis revealed that LC3, MDA, CAT and mTOR were strong predictors of PMI.  Conclusion: According to the study's findings, the levels of LC3 and mTOR expression in liver tissue can be utilized to calculate the PMI. LC3 was a strong independent indicator for PMI estimation.
Keywords
post mortem interval; LC3; Liver; aluminum phosphide toxicity; mTOR; MDA; CAT
References
 

  1. Montanari,, Giorgetti, R., Busardò, F. P., Giorgetti, A., Tagliabracci, A., & Alessandrini, F. (2021). Suitability of miRNA assessment in postmortem interval estimation. European Review for Medical and Pharmacological Sciences, 25(4), 1774-1787.‏
  2. Sangwan, A., Singh, S. P., Singh, P., Gupta, O. P., Manas, A., & Gupta, S. (2021). Role of molecular techniques in PMI estimation: An update. Journal of Forensic and Legal Medicine, 83, 102251.‏
  3. Thakral,, Purohit, P., Modi, A., Mishra, R., Sinha, A., & Setia, P. (2024). Autophagy genes (HMGB1, Bcl2, Bax) fold change expression in cadaveric heart tissue to estimate time since death: A unique approach. Archives of Legal Medicine, 15(1), 100423.‏
  4. Ahmad, A., Ghaleb, S. S., Zaki, A. R., Kamel, A. O., & Eid, A. (2024). The role of miRNA-21 and hypoxia inducible factor-1 in predicting post mortem interval in cardiac muscles of aluminum phosphide deaths. Journal of Forensic and Legal Medicine, 106, 102726.‏
  5. Ohkawa, H., Ohishi, N., & Yagi, K. (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical biochemistry, 95(2), 351-358.‏
  6. Aebi, H. (1984). [13] Catalase in vitro. In Methods in enzymology(Vol. 105, pp. 121-126). Academic press.‏
  7. Zhang, Y., Jing, Y., Qiao, J., Luan, B., Wang, X., Wang, L., & Song, Z. (2017). Activation of the mTOR signaling pathway is required for asthma onset. Scientific reports, 7(1), 4532.‏
  8. Yu, W., Liao, J., Yang, F., Zhang, H., Chang, X., Yang, Y., ... & Tang, Z. (2021). Chronic tribasic copper chloride exposure induces rat liver damage by disrupting the mitophagy and apoptosis pathways. Ecotoxicology and environmental safety, 212, 111968.‏
  9. Bancroft TD, Stevens A (1996) Turner DR. theory and practice of histological technique, 4th edition. Churchill, Livingston, New York, London, San Francisco, Tokyo.
  10. Yahia, , El-Amir, Y. O., & Sadek, A. A. I. (2018). Early postmortem biochemical and histopathological changes in the kidney, liver, and muscles of dogs. Comparative Clinical Pathology, 27, 1447-1455.‏
  11. Chan YH (2003a): Biostatistics102: Quantitative Data – Parametric & Non-parametric Tests. Singapore Med J.;44(8): 391-396.
  12. Chan YH (2003b): Biostatistics 104: Correlational Analysis. Singapore Med J.;44(12): 614-619.
  13. Chn YH (2004): Biostatistics 201: linear regression analysis. Singapore Med J.;45(2): 55-61.
  14. Maiese, , Scatena, A., Costantino, A., Di Paolo, M., La Russa, R., Turillazzi, E., ... & Fineschi, V. (2021). MicroRNAs as useful tools to estimate time since death. A systematic review of current literature. Diagnostics, 11(1), 64.‏
  15. Zapico, S. C., & Adserias-Garriga, J. (2022). Postmortem interval estimation: new approaches by the analysis of human tissues and microbial communities’ changes. Forensic Sciences, 2(1), 163-174.‏
  16. Martínez, P. N., Menéndez, S. T., de los Ángeles Villaronga, M., Ubelaker, D. H., García-Pedrero, J. M., & Zapico, S. C. (2019). The big sleep: Elucidating the sequence of events in the first hours of death to determine the postmortem interval. Science & Justice, 59(4), 418-424.‏
  17. Aghebat-Bekheir, S., & Abdollahi, M. (2024). Discovering the most impactful treatments for aluminum phosphide cardiotoxicity gleaned from systematic review of animal studies. Human & Experimental Toxicology, 43, 09603271241290922.‏
  18. Khezri, M. R., Varzandeh, R., & Ghasemnejad-Berenji, M. (2022). Protective effects of metformin against aluminum phosphide-induced acute hepato-renal damage in rats: an experimental approach. Pesticide Biochemistry and Physiology, 188, 105258.‏
  19. Sener, M. T., Suleyman, H., Hacimuftuoglu, A., Polat, B., Cetin, N., Suleyman, B., & Akcay, F. (2012). Estimating the postmortem interval by the difference between oxidant/antioxidant parameters in liver tissue. Advances in Clinical and Experimental Medicine: Official Organ Wroclaw Medical University, 21(6), 727-733.‏
  20. Shaaban, A. A. E. R., Farrag, D. I. M., Bayoumy, A., & Saad, E. (2017). Estimation of early postmortem interval by biochemical changes in brain and liver of rats using some oxidant and antioxidant parameters. The Egyptian Journal of Forensic Sciences and Applied Toxicology, 17(1), 147-162.‏
  21. El-Noor, M. M. A., Elhosary, N. M., Khedr, N. F., & El-Desouky, K. I. (2016). Estimation of early postmortem interval through biochemical and pathological changes in rat heart and kidney. The American journal of forensic medicine and pathology, 37(1), 40-46.‏
  22. Welson, N. N., Gaber, S. S., Batiha, G. E. S., & Ahmed, S. M. (2021). Evaluation of time passed since death by examination of oxidative stress markers, histopathological, and molecular changes of major organs in male albino rats. International Journal of Legal Medicine, 135, 269-280.‏
  23. Paltian, J., da Fonseca, C. A., Pinz, M. P., Luchese, C., & Antunes Wilhelm, E. (2019). Post-mortem interval estimative through determination of catalase and Δ-aminolevulinate dehydratase activities in hepatic, renal, skeletal muscle and cerebral tissues of Swiss mice. Biomarkers, 24(5), 478-483.‏
  24. Öztürk, C., Şener, M. T., Şener, E., Yılmaz, İ., Akcay, F., & Süleyman, H. (2013). The investigation of damage in the muscle tissue with the oxidant/antioxidant balance and the extent of postmortem DNA damage in rats.
  25. Zapico, S., Menéndez, S. T., & Núñez, P. (2014). Cell death proteins as markers of early postmortem interval. Cellular and molecular life sciences, 71, 2957-2962.
  26. Wan, L., Wang, Y., Li, J., Wang, Y., & Zhang, H. (2022). Inhibition of the AKT/mTOR pathway negatively regulates PTEN expression via miRNAs: Effect of Akt/mTOR inhibition on PTEN expression. Acta Biochimica et Biophysica Sinica, 54(11), 1637.‏
  27. Booth, A., Tavallai, S., Hamed, H. A., Cruickshanks, N., & Dent, P. (2014). The role of cell signalling in the crosstalk between autophagy and apoptosis. Cellular signalling, 26(3), 549-555.‏
  28. Mostafa, H. E. S., El-Shafei, D. A., Abouhashem, N. S., & Alaa El-Din, E. A. (2023). Could skeletal muscle changes provide a reliable method for estimating the time since death: A histological, biochemical, and DNA study. Australian Journal of Forensic Sciences, 55(1), 46-58.
 

 

Statistics
Article View: 7
PDF Download: 5