GYPA、CD68和TNF-α蛋白鉴别水中晚期尸体生前死后伤的作用研究  

A study on the role of GYPA,CD68 and TNF-α proteins in the identification of antemortem and postmortem injuries in late-stage submersion corpses

作　　者：向明亮 王廷宏 刘勤进 王陶 罗涛[1] 张伟[1] 章丽霞[1] 邓小冬[1] 刘云[1,2] Xiang Mingliang;Wang Tinghong;Liu QinJin;Wang Tao;Luo Tao;Zhang Wei;Zhang Lixia;Deng Xiaodong;Liu Yun(Department of Forensic Pathology,School of Basic Medical Science&Forensic Medicine,North Sichuan Medical College,Nanchong,637000 China;Sichuan Key Laboratory of Medical Imaging,Affiliated Hospital of North Sichuan Medical College,Nanchong,637000 China)

机构地区：[1]川北医学院基础医学与法医学院法医病理教研室,四川南充637000 [2]川北医学院医学影像四川省重点实验室,四川南充637000

出　　处：《中国法医学杂志》2024年第2期196-202,共7页Chinese Journal of Forensic Medicine

基　　金：四川省自然科学基金(2022NSFSC1286);南充市市校科技战略合作专项(20SXQT0327、22SXZRKX0004、22SXQT0177);四川省基层卫生事业发展研究中心资助项目(SWFZ19-Z-56)。

摘　　要：目的探讨血型糖蛋白A(Glycophorin A,GYPA)、CD68和肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)蛋白在水中晚期尸体生前伤和死后伤的表达情况,为在水中晚期生前死后伤鉴定筛选标志物。方法18只SD大鼠分为对照组(n=6)、生前挫伤浸没组(n=6)和死后挫伤浸没组(n=6),生前挫伤浸没组采用机械失重技术建立SD大鼠右后肢挫伤模型,死后挫伤浸没组是SD大鼠处死后用机械失重技术建立右后肢挫伤模型,对照组不予致伤处理,所有大鼠均浸没72 h。肉眼观察及HE染色观察生前伤和死后伤的情况;免疫组织化学染色(Immunohistochemistry,IHC)和蛋白质免疫印迹(Western blot)检测GYPA、CD68及TNF-α蛋白在挫伤组织表达情况;随后在实际案例中予以验证。结果肉眼观察和HE染色均无法较好地鉴别浸没72 h大鼠的生前伤和死后伤。免疫组织化学染色发现生前挫伤浸没组软组织GYPA表达呈阳性,CD68在炎性细胞膜呈阳性表达,TNF-α在胞质表达呈弱阳性,GYPA、CD68和TNF-α蛋白表达均显著高于死后挫伤浸没组和对照组(P<0.05)。GYPA、CD68和TNF-α蛋白在鉴别浸没尸体生前伤和死后伤实际案例中亦有相似结果。结论GYPA、CD68和TNF-α蛋白有望成为水中浸没尸体生前伤和死后伤鉴别的分子指标之一。Objective To investigate the expression of Glycophorin A(GYPA),CD68 and Tumor necrosis factor-α(TNF-α)proteins in the antemortem and postmortem injuries in late-stage submersion,and to identify screening markers for identification of antemortem and postmortem injuries in late-stage submersion.Methods Eighteen SD rats were divided into control group(n=6),antemortem contusion submersion group(n=6)and postmortem contusion submersion group(n=6).In the antemortem contusion submersion group and postmortem contusion submersion group,the right hind limb contusion models were established by the mechanical weightlessness technique in the alive rats and dead rats respectively.All rats were submerged in river water for 72 hours.The injury characteristics were evaluated by gross observation and HE staining.The expression of GYPA,CD68 and TNF-α proteins in the contusion tissues were assessed by immunohistochemistry(IHC)and Western Blot(WB).Subsequently,the changes in the expression of GYPA,CD68 and TNF-α were verified in a real case.Results Gross observation and HE staining could not distinguish the antemortem and postmortem injuries in the rats submerged in water for 72h.IHC analysis showed that the GYPA and CD68 expression was positive on the inflammatory cell membrane,and the TNF-α expression was weakly positive in the cytoplasm.The protein expression levels of GYPA,CD68 and TNF-α in the antemortem contusion immersion group were significantly higher than those in the postmortem contusion immersion group and in the control group(P<0.05).Moreover,the antemortem and postmortem injuries in a real case were assessed by detection of GYPA,CD68,and TNF-α protein expression by IHC.Conclusion GYPA,CD68 and TNF-α are expected to be the molecular markers for the identification of antemortem and postmortem injuries in submerged bodies.

关 键 词：法医病理学 生前伤 死后伤 血型糖蛋白A 肿瘤坏死因子-α CD68 

分 类 号：D919.6[医药卫生—法医学]