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通过检测雄性白化大鼠主要器官的氧化应激标志物、组织病理学和分子变化来评估死亡时间。

Evaluation of time passed since death by examination of oxidative stress markers, histopathological, and molecular changes of major organs in male albino rats.

机构信息

Department of Forensic Medicine and Clinical Toxicology, Faculty of Medicine, Beni-Suef University, Beni-Suef, Egypt.

Department of Biochemistry, Faculty of Medicine, Minia University, Minia, Egypt.

出版信息

Int J Legal Med. 2021 Jan;135(1):269-280. doi: 10.1007/s00414-020-02463-1. Epub 2020 Nov 25.

Abstract

Recent biochemical, metabolic, and molecular profiles of various body fluids showed more accurate correlation to the postmortem interval than the traditional physical examination. Our study aimed to evaluate time passed since death in relation to oxidative stress markers, HMGB1 genetic expression, histopathological examination, and BCL2 immunohistochemical analysis in major organs (heart, kidney, and testis). Forty-two adult male rats were included and randomly divided into seven equal groups. After sacrification, the rodents were kept at room temperature and major organs were obtained at 0, 12, 24, 48, 72, 96, and 120 h. Malonaldehyde (MDA), superoxide dismutase (SOD), reduced glutathione (GSH) tissue levels, High mobility group box 1 protein (HMGB1) gene expression, histopathological, and B cell lymphoma 2 (BCL2) immunohistochemical expressions were analyzed. Postmortem interval was correlated to different tissue levels of MDA, SOD, and GSH. HMGB1 showed enhanced postmortem gene expression with a peak at 48 h after death. Obvious time-dependent histopathological changes were observed in all the examined organs. Dilated spaces, extravasation, and fragmentation scores in heart specimens were higher at 96 and 120 h compared with the other groups. Renal changes in the form of shrunken glomeruli, loss of tubular epithelium, and hyalinization and testicular findings in the form of epithelial detachment, vacuolation, and loss of sperms started at 72 h postmortem. BCL2 expression began to decrease 24 h and became negative at 96 h after death. In conclusion, HMGB1 gene expression can be used for estimation of time passed since death as it shows time-dependent changes in the form of a progressive increase with a peak at 48 h then it begins to decline. Oxidants and antioxidants are correlated to PMI until 120 h after death. Histopathological changes in the heart, kidney, and testis are also time-dependent until the 5th day after death. BCL2 immunohistochemical expression begins to decline 24 h until 96 h after death when it becomes negative.

摘要

最近,各种体液的生化、代谢和分子特征显示出与传统的体格检查相比,与死后时间间隔有更准确的相关性。我们的研究旨在评估与氧化应激标志物、HMGB1 基因表达、组织病理学检查和 BCL2 免疫组织化学分析相关的死后时间间隔,这些标志物分别在主要器官(心脏、肾脏和睾丸)中进行检测。共纳入 42 只成年雄性大鼠,并随机分为 7 组。处死动物后,将啮齿动物在室温下保存,并在 0、12、24、48、72、96 和 120 小时后获取主要器官。分析丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)组织水平、高迁移率族蛋白 B1(HMGB1)基因表达、组织病理学和 B 细胞淋巴瘤 2(BCL2)免疫组织化学表达。死后时间间隔与不同组织水平的 MDA、SOD 和 GSH 呈正相关。HMGB1 表现出增强的死后基因表达,在死后 48 小时达到峰值。所有检查器官均观察到明显的时间依赖性组织病理学变化。心脏标本的扩张空间、渗出和碎裂评分在 96 和 120 小时时高于其他组。72 小时后出现肾小球缩小、管状上皮丧失、玻璃样变等肾脏改变和上皮脱落、空泡形成和精子丧失等睾丸发现。BCL2 表达在死后 24 小时开始下降,96 小时后变为阴性。总之,HMGB1 基因表达可用于估计死后时间,因为它表现出时间依赖性变化,呈逐渐增加的形式,在 48 小时达到峰值,然后开始下降。氧化剂和抗氧化剂与死后 120 小时内的 PMI 相关。心脏、肾脏和睾丸的组织病理学变化也一直持续到死后第 5 天。BCL2 免疫组织化学表达在死后 24 小时开始下降,96 小时后变为阴性。

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