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Forensic diagnosis of fatal hypothermia is considered difficult because there are no specific findings. Accordingly, exploration of novel fatal hypothermia-specific findings is important. To elucidate the molecular mechanism of homeostasis in hypothermia and identify novel molecular markers to inform the diagnosis of fatal hypothermia, we focused on microRNA expression in skeletal muscle, which plays a role in cold-induced thermogenesis in mammals. We generated rat models of mild, moderate, and severe hypothermia, and performed body temperature-dependent microRNA expression analysis of the iliopsoas muscle using microarray and quantitative real-time PCR (qRT-PCR). The results show that  rno - miR-374-5p  expression was significantly induced only by severe hypothermia. Luciferase reporter assay and qRT-PCR results indicated that  Mex3B  expression was regulated by  rno-miR-374-5p  and decreased with decreasing body temperature. Gene ontology analysis indicated the involvement of  Mex3B  in positive regulation of GTPase activity. siRNA analysis showed that  Mex3B  directly or indirectly regulated  Kras  expression in vitro ,  and significantly changed the expression of apoptosis-related genes and proteins. Collectively, these results indicate that  rno-miR-374-5p  was activated by a decrease in body temperature, whereby it contributed to cell survival by suppressing  Mex3B  and activating or inactivating  Kras . Thus,  rno-miR-374-5p  is a potential supporting marker for the diagnosis of fatal hypothermia.

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Body temperature-dependent microRNA expression analysis in rats: rno-miR-374-5p regulates apoptosis in skeletal muscle cells via Mex3B under hypothermia