Determination of Postmortem Interval Using the Oxidative Stress Markers, Histopathological, Immunohistochemical, and Molecular Changes of Spleen, Pancreas, and Heart: Experimental study
International Journal of Medical Toxicology and Forensic Medicine,
Vol. 16 (2026),
1 January 2026
,
Page 1-13
https://doi.org/10.22037/ijmtfm.v16.50951
Abstract
Background: A cornerstone of forensic practice is determining the postmortem interval to resolve suspicious deaths and assist in criminal investigations. The work aims to estimate the postmortem interval (PMI) using oxidative stress markers, and to Assess Molecular, histopathological, and immunohistochemical changes in the spleen, pancreas, and heart of adult albino rats.
Methods: 30 adult albino rats were divided into five groups. All rats were sacrificed and kept at room temperature. Dissection of the spleen, pancreas, and heart was done at 0 (control), 6, 24, 48, and 72 hours. The following parameters were measured: malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD), hypoxia-associated factor (HAF), apoptosis-inducing factor (AIF), and B-cell lymphoma 2 (BCL2) immunostaining.
Results: MDA, GSH, and SOD showed strong positive correlations with PMI in the pancreas, whereas in the heart, GSH showed a fair positive correlation with PMI.SOD showed a strong positive correlation with PMI in the spleen, while GSH showed a strong negative correlation with PMI. HAF showed a strongly significant positive association with PMI in the heart and the spleen, whereas in the pancreas, it showed a fair correlation. AIF showed a significant negative correlation with PMI in the heart. Histological changes showed time-dependent changes. BCL2 immunohistochemical expression showed a significant decrease over time in all organs.
Conclusion: Oxidative stress markers, Molecular, histopathological, and immunohistochemical changes can help estimate PMI. Four novel equations were developed to estimate PMI using the studied parameters across different organs.
- Postmortem interval, BCL2, Molecular changes, Oxidative stress
How to Cite
References
[1] Thakral S, Purohit P, Mishra R, Gupta V, Setia P. The impact of RNA stability and degradation in different tissues to the determination of postmortem interval: A systematic review. Forensic Sci Int. 2023;111772. [DOI: 10.1016/j.forsciint.2023.111772]
[2] Elghamry HA, Hassan FM, Mohamed MI, Abdelfattah DS, Abdelaal AG. Estimation of the postmortem interval using GAPDH mRNA in skin and heart tissues of albino rats at different environmental conditions. Egypt J Forensic Sci. 2018;8:1-6. [DOI: 10.1186/s41935-018-0059-3]
[3] Brooks JW. Postmortem changes in animal carcasses and estimation of the postmortem interval. Vet Pathol. 2016;53(5):929-40. [DOI: 10.1177/0300985816629720]
[4] Lv YH, Ma KJ, Zhang H, He M, Zhang P, Shen YW, et al. A time course study demonstrating mRNA, microRNA, 18S rRNA, and U6 snRNA changes to estimate PMI in deceased rat spleen. J Forensic Sci. 2014;59(5):1286-94. [DOI: 10.1111/1556-4029.12470]
[5] Pasaribu RS, Auerkari EI, Suhartono AW. Histological changes in oral mucosa (gingiva) as a method for estimating postmortem interval: A literature review. Saudi Dent J. 2024. [DOI: 10.1016/j.sdentj.2024.05.002]
[6] El-Noor MM, Elhosary NM, Khedr NF, El-Desouky KI. Estimation of early postmortem interval through biochemical and pathological changes in rat heart and kidney. Am J Forensic Med Pathol. 2016;37(1):40-6. [DOI: 10.1097/PAF.0000000000000213]
[7] Kiernan JA. Staining, histochemistry and histotechnology FAQ. [Link]
[8] Suvarna KS, Layton C, Bancroft JD. Bancroft’s theory and practice of histological techniques. 8th ed. Elsevier; 2018. [DOI: 10.1016/C2015-0-00143-5]
[9] Chan YH. Biostatistics 102: Quantitative data – parametric & non-parametric tests. Singapore Med J. 2003;44(8):391-6. [Link]
[10] Chan YH. Biostatistics 104: Correlational analysis. Singapore Med J. 2003;44(12):614-9. [Link]
[11] Chan YH. Biostatistics 201: Linear regression analysis. Singapore Med J. 2004;45(2):55-61. [Link]
[12] Zaki AR, Tohamy AF, Yaseen NE. Estimation of postmortem intervals by some biochemical changes and DNA degradation in rat brain and skeletal muscle tissues. Mansoura J Forensic Med Clin Toxicol. 2017;25(1):59-78. [Link]
[13] Welson NN, Gaber SS, Batiha GE, Ahmed SM. Evaluation of time passed since death by examination of oxidative stress markers, histopathological, and molecular changes of major organs in male albino rats. Int J Legal Med. 2021;135:269-80. [DOI: 10.1007/s00414-020-02428-4]
[14] Shaaban DA, Farrag DI, Bayoumy A, Saad E. Estimation of early postmortem interval by biochemical changes in brain and liver of rats using some oxidant and antioxidant parameters. Egypt J Forensic Sci Appl Toxicol. 2017;17(1):147-62. [Link]
[15] Ali D, Ahmed M. Prostatic estimation of time passed since death: Oxidative stress markers, histological and immunohistochemical examination (an experimental study). Ain Shams J Forensic Med Clin Toxicol. 2023;40(1):22-33. [DOI: 10.21608/ajfm.2023.295402]
[16] Mostafa HE, El-Shafei DA, Abouhashem NS, Alaa El-Din EA. Could skeletal muscle changes provide a reliable method for estimating the time since death: A histological, biochemical, and DNA study. Aust J Forensic Sci. 2023;55(1):46-58. [DOI: 10.1080/00450618.2021.1964975]
[17] Hegazy AM, Nasr SM, Aziem SH. Ultrastructure of cerebral cortex investigation during early postmortem changes in a rat model. Indian J Forensic Med Pathol. 2020;13(3):210-8. [Link]
[18] da Fonseca CA, Paltian J, Dos Reis AS, Bortolatto CF, Wilhelm EA, Luchese C. Na+/K+-ATPase, acetylcholinesterase and glutathione S-transferase activities as new markers of postmortem interval in Swiss mice. Leg Med (Tokyo). 2019;36:67-72. [DOI: 10.1016/j.legalmed.2018.11.005]
[19] Su SB, Motoo Y, Xie MJ, Mouri H, Asayama K, Sawabu N. Superoxide dismutase is induced during rat pancreatic acinar cell injury. Pancreas. 2002;24(2):146-52. [DOI: 10.1097/00006676-200203000-00008]
[20] Estaras M, Martinez-Morcillo S, García A, Martinez R, Estevez M, Perez-Lopez M, et al. Pancreatic stellate cells exhibit adaptation to oxidative stress evoked by hypoxia. Biol Cell. 2020;112(10):280-99. [DOI: 10.1111/boc.201900073]
[21] Zheng W, Huang LZ, Zhao L, Wang B, Xu HB, Wang GY, et al. Superoxide dismutase activity and malondialdehyde level in plasma and morphological evaluation of acute severe hemorrhagic shock in rats. Am J Emerg Med. 2008;26(1):54-8. [DOI: 10.1016/j.ajem.2007.02.012]
[22] Peng D, Lv M, Li Z, Tian H, Qu S, Jin B, et al. Postmortem interval determination using mRNA markers and DNA normalization. Int J Legal Med. 2020;134:149-57. [DOI: 10.1007/s00414-019-02116-3]
[23] Bai X, Peng D, Li Z, Tian H, Zhang L, Yang D, et al. Postmortem interval (PMI) determination by profiling of HAF mRNA degradation using RT-qPCR. Forensic Sci Int Genet Suppl Ser. 2017;6:182-3. [DOI: 10.1016/j.fsigss.2017.09.073]
[24] Öztürk C, Şener MT, Şener E, Yılmaz İ, Akcay F, Süleyman H. The investigation of damage in muscle tissue with oxidant/antioxidant balance and the extent of postmortem DNA damage in rats. Life Sci J. 2013;10(3):1631-7. [Link]
[25] Tomita Y, Nihira M, Ohno Y, Sato S. Ultrastructural changes during in situ early postmortem autolysis in kidney, pancreas, liver, heart and skeletal muscle of rats. Leg Med (Tokyo). 2004;6(1):25-31. [DOI: 10.1016/j.legalmed.2003.08.002]
[26] Mohamed A, Elbohi K, Sharkawy N, Hassan M. Biochemical and apoptotic biomarkers as indicators of time elapsed since death in experimentally induced traumatic brain injury. SM J Forensic Res Criminol. 2017;1(2):1-7. [Link]
[27] Elias EZ, Osman K, Aziz SM, Mohamed J, Mansar AH, Ibrahim SF. Determination of time of death based on basic histological stain and immunostain changes. J Sains Kesihat Malays. 2004;2:63-70. [Link]
[28] Khalifa FN, Hosny SA, Moawad AM. Histobiochemical changes in early postmortem interval in liver, pancreas, skin and kidney of adult male albino rats. Rechtsmedizin. 2022;32(5):374-85. [DOI: 10.1007/s00194-022-00554-2]
- Abstract Viewed: 49 times
- PDF Downloaded: 51 times