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肺细小支气管形态及淋巴结PHH3表达变化推断死亡时间

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目的 研究大鼠细小支气管及淋巴结形态学变化用于推断早期死亡时间(Postmortem Interval,PMI)的可行性.方法 35 只 8 周龄雄性SD大鼠以颈椎脱臼法处死,置于恒温恒湿箱内,分别于死后 24 h内各时间点取 5只提取肺组织行常规HE及磷酸化组蛋白H3(Phospho-Histone H3,PHH3)免疫组织化学染色,使用Image J 分别测量粘膜层面积(S粘膜)、粘膜层基底侧与平滑肌层管腔侧之间面积(S粘膜外)及管腔面积(S管腔),并计算S粘膜/S管腔(粘膜占比)、S粘膜外/S管腔(粘膜外占比)及S粘膜外/S粘膜;使用Image J IHC profiler测量淋巴结细胞核PHH3 免疫组织化学染色阳性程度,使用Graphpad Prism 9.5 对上述计算结果及PHH3 染色阳性程度进行统计学分析.结果 粘膜占比随PMI增加呈下降趋势(0 h与18 h及24 h相比具有统计学差异,P<0.05);粘膜外占比及S粘膜外/S粘膜随PMI增加呈上升趋势(0 h与其余各时间点相比均具有统计学差异,P<0.05);PHH3 免疫组织化学染色阳性程度在死后24 h明显减弱(P<0.05).结论 肺细小支气管、淋巴结形态学变化在死后24 h内呈现一定的规律性,有助于推断早期PMI,淋巴结细胞核PHH3有望成为鉴别早期与晚期PMI的临界指标.
Study on the morphological changes of pulmonary bronchioles and the expression of PHH3 in lymph node nuclei to infer the early time of death
Objective To study the feasibility of using morphological changes of bronchioles and lymph nodes in rats for estimating the postmortem interval(PMI).Methods Thirty-five 8-week-old male SD rats were sacrificed by cervical dislocation and placed in a constant temperature and humidity box.Lung tissues of 5 rats were extracted at each time point within 24 hours after death for routine HE and PHH3 immunohistochemical staining.Image J was used to measure mucosal area,extramucosal area,luminal area of small bronchi,Then calculate the ratio of mucosal area to luminal area,extramucosal area to extramucosal,extramucosal area to mucosal area.imageJIHCprofiler was used to measure the positive degree of PHH3 immunohistochemical staining in lymph node nuclei,and the above calculation results and the positive degree of PHH3 staining were statistically analyzed by GraphPad Prism 9.5.Results The ratio of mucosal area to lumen area showed an overall downward trend with the increase of PMI(0h was statistically different compared with 18h and 24h,P<0.05).The ratio of extramucosal to lumen area and extramucosal to mucosa area showed an upward trend with the increase of PMI(0h was statistically different from other time points,P<0.05),the positive degree of PHH3 immunohistochemical staining was significantly decreased at 24 h after death(P<0.05).Conclusion The morphological changes of pulmonary bronchiole and lymph nodes showed a certain regularity within 24h after death,which was helpful to infer the early PMI,lymph node nucleus PHH3 was expected to be a critical index to distinguish early and late PMI.

Forensic pathologyPostmortem intervalHistomorphologySmall bronchiLymph nodesPhospho-Histone H3(PHH3)

张永昊、李明明、何梁超、陈刚、王荣帅、陈光、吴茂旺

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皖南医学院法医学院,安徽 芜湖 241000

芜湖知微病理诊断中心,安徽 芜湖 241000

法医病理学 死亡时间 组织形态学 细小支气管 淋巴结 磷酸化组蛋白H3(PHH3)

2024

中国法医学杂志
中国法医学会

中国法医学杂志

CSTPCD
影响因子:0.352
ISSN:1001-5728
年,卷(期):2024.39(6)