Cell-Biological Study on Medicolegal Diagnosis of Injury

损伤法医学诊断的细胞生物学研究

• 批准号: 07457118
• 负责人: TSUJI Tsutomu
• 金额: $ 2.62万
• 依托单位: Wakayama Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457118/
• 关键词: Injury; Cell Death; Apoptosis; Necrosis; TNF alpha; TNF-α; 細胞死機構

In this study, we investigated cell-biologically the cell death generating by injuries, on the basis of two mechanisms of the cell death, these are necrosis and apoptosis.To heat the liver of rts, a heat device having a built-in PID electron regulater system was newly developed by us. The liver of rats laparotomized under anesthesia was heated at 37ﾟC,50ﾟC and 65ﾟC for 5 min. with a heated needle connected to the device, and thereafter the rats were survived with in the given time. Appearance of the cell death in the tissue injured by heat was analyzed, in progress of the survival time, by means of staining of the tissue with hematoxylin-eosin (HE) and TUNEL method and agarosegel electrophoresis of DNA extracted from the tissue injured.Consequently, apoptosis appeared, in all heat temperatures, with bimodal pattern showing peak at about 2 and 10 hours after heat. Apoptosis in the first peak dottedly appeared, in heat temperature at 37ﾟC,in the whole region affected by heat. When the li … More ver was heated at 50ﾟC and 65ﾟC,it first appeared on outer edge in the region affected by heat, and then made progress to the center of the region, condensation of chromatin was, in HE staining, remarkable at border between the injured and normal tissues. These findings were different from those in 37ﾟC.It seems that apoptosis inthe first peak is caused by direct action of temperature. Apoptosis in the second peak, in all heat temperature, made progress from outer edge of the injured region to the center, time course of progression of the apoptosis was almost concurrent with that of infiltration of neutrophil to the injured region. Ladder formation was observed, when DNA extracted from the tissue showing apoptosis was applied for agarosegel electrophoresis. Necrosis in the injured tissue appeared in the same pattern with appearance of apoptosis in time course and on location, necrosis and apoptosis were observed in almost same degree up to appearance of the second peak of apoptosis. After 24 hours of heat, apoptosis cells were phagocytosed, only necrotic tissue remained. It seems that appearance of necrosis and apoptosis is caused by action of TNF alpha derived from neutrophils, since generation of TNF alpha was confirmed electron microscopically.In this study, it became cell-biologically evident that apoptosis is intensely concerned with cell death in the injury tissue. Less

在本研究中，我们从细胞生物学角度研究了损伤引起的细胞死亡，基于细胞死亡的两种机制，即坏死和凋亡。为了加热 rts 的肝脏，使用了内置 PID 电子的加热装置调节器系统是我们新开发的，将麻醉下剖腹的大鼠肝脏用连接到该装置的加热针加热到37°C、50°C和65°C。此后，在给定时间内使大鼠存活，通过用苏木精-伊红(HE)和TUNEL方法对组织进行染色，分析受热损伤的组织中细胞死亡的情况。对从受伤组织中提取的DNA进行琼脂糖凝胶电泳。因此，在所有加热温度下都出现了细胞凋亡，并且双峰模式显示在加热后约2小时和10小时出现峰值，第一个峰值出现细胞凋亡。在加热温度为37°C时，在整个受热区域中出现点状。当将li … More ver加热到50°C和65°C时，它首先出现在受热区域的外边缘，并且。然后进展到该区域的中心，在HE染色中，在受伤组织和正常组织之间的边界处染色质的浓缩是显着的，这些结果与37℃下的结果不同。看来第一个峰值的细胞凋亡是由温度的直接作用引起的，在所有加热温度中，细胞凋亡都是从损伤区域的外缘向中心进行的，细胞凋亡的进展时间进程几乎与同时进行。当从显示细胞凋亡的组织中提取的DNA进行琼脂糖凝胶电泳时，观察到中性粒细胞浸润到损伤区域。在时间和位置上，损伤组织中出现与细胞凋亡相同的模式。在加热24小时后，观察到几乎相同程度的坏死和凋亡出现。加热24小时后，凋亡细胞被吞噬，仅保留坏死组织，看来坏死和凋亡的出现是由TNFα的作用引起的。源自中性粒细胞，因为 TNF α 的产生已被电子显微镜证实。在这项研究中，细胞生物学证据表明细胞凋亡与损伤组织中的细胞死亡密切相关。