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Background <p>Heatstroke and fever-related deaths pose significant diagnostic challenges in forensic practice due to overlapping clinical manifestations. This study aims to identify specific molecular markers distinguishing these conditions through transcriptomic profiling of liver tissues in mouse models.</p> Methods <p>We established mouse models of severe heatstroke (exposure to 42℃ high-temperature environment) and fever (LPS-induced), and collected liver tissues for RNA sequencing. Differentially expressed genes (DEGs) were identified by DESeq2 and analyzed via GO and KEGG enrichment. Five candidate genes (<i>Hspa1a</i>,<i> Hspa1b</i>,<i> Cyp7a1</i>,<i> Arrdc3</i>, and <i>G6pc</i>) were validated by RT-qPCR and Western blotting.</p> Results <p>Transcriptome profiling revealed 5,567 DEGs between the two groups, including 142 heatstroke-specific and 254 fever-specific genes. Heatstroke was characterized by significant up-regulation of <i>Hspa1a</i>,<i> Hspa1b</i>,<i> Cyp7a1</i>,<i> Arrdc3</i>, and <i>G6pc</i>, implicating pathways related to protein homeostasis, glucose metabolism, and energy regulation. In contrast, fever predominantly induced immune- and inflammation-related genes such as <i>Gbp2</i>,<i> Lcn2</i>,<i> Gm12250</i>,<i> Zbp1</i>, and <i>Ccrl2</i>. Importantly, RT-qPCR and Western blot assays consistently validated the differential expression of the five key genes (<i>Hspa1a</i>,<i> Hspa1b</i>,<i> Cyp7a1</i>,<i> Arrdc3</i>, and <i>G6pc</i>), confirming their potential as reliable biomarkers for distinguishing heatstroke from fever.</p> Conclusion <p>This study provides comparative transcriptomic evidence distinguishing heatstroke from fever. The identified markers, particularly <i>Hspa1a</i>,<i> Hspa1b</i>,<i> Cyp7a1</i>,<i> Arrdc3</i>, and <i>G6pc</i>, may serve as potential forensic indicators for differentiating hyperthermia-related deaths, while also offering new insights into the molecular responses underlying heat stress and fever.</p>

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Identification of specific molecular markers in severe heatstroke using transcriptomics techniques

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年10月13日

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  • Zhiao Duan,
  • Mei Wang,
  • Yunchao Zhou,
  • Jianghuan Lu,
  • Yixin Ma,
  • Yuzhao Xu,
  • Jianqiang Deng,
  • Hui Liu,
  • Jianhua Chen

摘要

Background

Heatstroke and fever-related deaths pose significant diagnostic challenges in forensic practice due to overlapping clinical manifestations. This study aims to identify specific molecular markers distinguishing these conditions through transcriptomic profiling of liver tissues in mouse models.

Methods

We established mouse models of severe heatstroke (exposure to 42℃ high-temperature environment) and fever (LPS-induced), and collected liver tissues for RNA sequencing. Differentially expressed genes (DEGs) were identified by DESeq2 and analyzed via GO and KEGG enrichment. Five candidate genes (Hspa1a, Hspa1b, Cyp7a1, Arrdc3, and G6pc) were validated by RT-qPCR and Western blotting.

Results

Transcriptome profiling revealed 5,567 DEGs between the two groups, including 142 heatstroke-specific and 254 fever-specific genes. Heatstroke was characterized by significant up-regulation of Hspa1a, Hspa1b, Cyp7a1, Arrdc3, and G6pc, implicating pathways related to protein homeostasis, glucose metabolism, and energy regulation. In contrast, fever predominantly induced immune- and inflammation-related genes such as Gbp2, Lcn2, Gm12250, Zbp1, and Ccrl2. Importantly, RT-qPCR and Western blot assays consistently validated the differential expression of the five key genes (Hspa1a, Hspa1b, Cyp7a1, Arrdc3, and G6pc), confirming their potential as reliable biomarkers for distinguishing heatstroke from fever.

Conclusion

This study provides comparative transcriptomic evidence distinguishing heatstroke from fever. The identified markers, particularly Hspa1a, Hspa1b, Cyp7a1, Arrdc3, and G6pc, may serve as potential forensic indicators for differentiating hyperthermia-related deaths, while also offering new insights into the molecular responses underlying heat stress and fever.