Department of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang, Henan 471023, China; Department of Forensic Medicine, Faculty of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China.
Department of Basic Medicine, Chongqing College of Traditional Chinese Medicine, Chongqing 402760, China.
Leg Med (Tokyo). 2023 Nov;65:102302. doi: 10.1016/j.legalmed.2023.102302. Epub 2023 Jul 29.
Identification of mechanical asphyxia deaths without obvious injuries is a difficult problem for forensic medicine. This study aimed to identify molecular biological markers to predict death from mechanical asphyxia (smother). We established a smother model of mice by over the head with plastic bag tightly until the mice died and applied label-free proteomic technology to identify differentially expressed proteins (DEPs) in heart. A total of 3307 proteins were quantified, and a Fold Change (FC) > 1.2 (or <1/1.2) and Q value < 0.05 were considered as DEPs. Through comparative analysis, we identified 606 DEPs compared to the control group, comprising 219 upregulated and 387 downregulated proteins. Bioinformatics analysis (MCODE analysis) showed that the candidate proteins were mainly involved in regulation of ribosome function, myocardial contraction and calcium regulation, regulation of coagulation and regulation of mitochondrial oxidative respiration. Seven of these proteins were validated using parallel reaction monitoring (PRM), including fibrinogen alpha chain (FIBA), fibrinogen gamma chain (FIBG), Calsequestrin-2 (CASQ2), NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 11 (NDUAB), NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 3 (NDUA3), NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13 (NDUAD) and Rab7 (RAB7A). CASQ2 and FIBG were further validated by immunohistochemistry. In conclusion, our results may provide some auxiliary indices for identifying the death from mechanical asphyxia.
法医学中，识别无明显损伤的机械性窒息死亡是一个难题。本研究旨在寻找可预测机械性窒息（捂压）死亡的分子生物学标志物。我们通过用塑料袋紧紧捂住头部的方式建立了小鼠捂压模型，直至小鼠死亡，并应用无标记蛋白质组学技术鉴定心脏中的差异表达蛋白（DEPs）。共定量了 3307 种蛋白质，FC＞1.2（或＜1/1.2）且 Q 值＜0.05 被认为是 DEPs。通过比较分析，与对照组相比，我们鉴定出 606 个 DEPs，包括 219 个上调蛋白和 387 个下调蛋白。生物信息学分析（MCODE 分析）显示，候选蛋白主要参与核糖体功能调节、心肌收缩和钙调节、凝血调节和线粒体氧化呼吸调节。使用平行反应监测（PRM）验证了其中的 7 种蛋白质，包括纤维蛋白原α链（FIBA）、纤维蛋白原γ链（FIBG）、钙结合蛋白 2（CASQ2）、NADH 脱氢酶[泛醌]1α亚基 11（NDUAB）、NADH 脱氢酶[泛醌]1α亚基 3（NDUA3）、NADH 脱氢酶[泛醌]1α亚基 13（NDUAD）和 Rab7（RAB7A）。通过免疫组织化学进一步验证了 CASQ2 和 FIBG。总之，我们的结果可能为识别机械性窒息死亡提供一些辅助指标。
