Forensic science international · 2026
Temperature-dependent alterations in skeletal muscle during the postmortem interval: Structural, metabolic, and lncRNA expression changes.
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Zhang Jiaji, Chen Jiaqiu, Wu Xiaoyan, Zhong Yunlong, Li Shoutian
Аннотация
BACKGROUND: Estimation of the postmortem interval (PMI) remains a challenge in forensic practice. Although environmental temperature is known to influence postmortem biological processes, its effects on structural, metabolic, and transcriptional changes in skeletal muscle remain poorly characterized.
METHODS: Male C57 mice were euthanized and carcasses were maintained at 30 °C, 20 °C, or 4 °C. Skeletal muscle samples were collected at 0, 4, 8, 12, 24, and 36 h postmortem. Histological alterations were assessed by H&E staining. Adenylate energy status was evaluated by measuring ATP, ADP, and AMP using high-performance liquid chromatography, followed by calculation of energy charge and the ATP/ADP ratio. Autophagy-related markers were analyzed by Western blot, transmission electron microscopy, and immunofluorescence. Long non-coding RNA (lncRNA) expression profiles were examined by sequencing, and selected candidates were validated by quantitative PCR.
RESULTS: Skeletal muscle structure progressively deteriorated with increasing PMI, with changes occurring earlier at 30 °C and delayed at 4 °C. ATP levels declined postmortem in all groups, accompanied by increased AMP and reduced energy charge and ATP/ADP ratio. The decline in energy status was accelerated at higher temperature and was slowed at lower temperature. Autophagy-related markers showed early elevation followed by a later reduction, with peak changes occurring earlier at higher temperature and later at lower temperature. Transcriptomic analysis identified 1643 high-confidence lncRNAs. Differential expression patterns varied among temperature groups. Six candidate lncRNAs displayed distinct time-dependent expression trajectories under different temperature conditions, and their temporal trends were described using fourth-order polynomial regression (R2 = 0.87-0.92).
CONCLUSION: Postmortem skeletal muscle exhibits structural, metabolic, and transcriptional alterations that are influenced by environmental temperature. These temperature-dependent temporal patterns may provide molecular information relevant to PMI assessment.
METHODS: Male C57 mice were euthanized and carcasses were maintained at 30 °C, 20 °C, or 4 °C. Skeletal muscle samples were collected at 0, 4, 8, 12, 24, and 36 h postmortem. Histological alterations were assessed by H&E staining. Adenylate energy status was evaluated by measuring ATP, ADP, and AMP using high-performance liquid chromatography, followed by calculation of energy charge and the ATP/ADP ratio. Autophagy-related markers were analyzed by Western blot, transmission electron microscopy, and immunofluorescence. Long non-coding RNA (lncRNA) expression profiles were examined by sequencing, and selected candidates were validated by quantitative PCR.
RESULTS: Skeletal muscle structure progressively deteriorated with increasing PMI, with changes occurring earlier at 30 °C and delayed at 4 °C. ATP levels declined postmortem in all groups, accompanied by increased AMP and reduced energy charge and ATP/ADP ratio. The decline in energy status was accelerated at higher temperature and was slowed at lower temperature. Autophagy-related markers showed early elevation followed by a later reduction, with peak changes occurring earlier at higher temperature and later at lower temperature. Transcriptomic analysis identified 1643 high-confidence lncRNAs. Differential expression patterns varied among temperature groups. Six candidate lncRNAs displayed distinct time-dependent expression trajectories under different temperature conditions, and their temporal trends were described using fourth-order polynomial regression (R2 = 0.87-0.92).
CONCLUSION: Postmortem skeletal muscle exhibits structural, metabolic, and transcriptional alterations that are influenced by environmental temperature. These temperature-dependent temporal patterns may provide molecular information relevant to PMI assessment.