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[细胞光度形态学检查中的核溶解现象]

[Phenomenon of karyolysis in cytophotomorphometric examinations].

作者信息

Parafiniuk M

出版信息

Ann Acad Med Stetin. 1998;Suppl 43:1-87.

PMID:9857529
Abstract

Studies were performed on hepatocytes of ten rabbits' livers. The one-year-old animals were sacrificed by a blow in the suboccipital region. Under aseptic conditions each liver was divided into 9 parts. One of them was examined at once (zero time) the remaining parts were placed in sterile vessels lined with gauze moistened in Ringer's solution, which created an environment of 100% humidity. Autolysis was being conducted for 8 hours at temperature 37 degrees C, carrying out the investigations with hourly intervals. The cells for studying were obtained by preparing contact smears that stemmed from a fresh cross-section of the liver segment. The smears were fixed in formalin vapour 40% proceeding for 2 hours at temperature 21 degrees C, and subsequently in 96% ethanol for 0.5 hour. Feulgen's reaction was run in accordance with the routine principles by using the basic fuschin NI 42510 of Fischer Firm (USA). The morpho-photometric measurements of nuclei were taken by means of computed device "Morphoquant" of Carl Zeiss Firm, Jena, 1982. The following measurements of the cell nuclei were accomplished, namely: A. morphometric--surface, circumferential length, bulged surface, ratio of diameters, unstained surface; B. photometric--medium extinction, circumferential extinction, total extinction. The results of studies were documented by 95 diagrams. On the basis of measurements taken, it has been established that, during the 7 hour-long period of autolysis, nucleus of hepatocyte is subjected to dynamic transformations, whereas the plotted function does not follow a linear course. During the observation the circumference and the surface resemble a polynomial with three extremes (Fig. 12, 21, 39); similarly the mean extinction and circumference extinction (attention is attracted by its severe course as compared to surface and circumference) (Fig. 57, 66). At the time of experiment the differentiation of nuclei population undergoes changes in the direction of its uniformity which concerns, to greater or lesser degree, all the investigated features (compare figures of histograms). The isolated group of tetraploidal nuclei is characterised by occupying another place. It is defined by 3 variables in spatial arrangement (Fig. 76, 80). The analysis of correlation allowed for determination of the dynamic connections, between the studied variables, which seem to be specific for a given experimental time (Tab. 1, 2). On the ground of the performed studies 7 final conclusions have been formulated: 1. Autolysis of the nuclei of hepatocytes is non-linear phenomenon; (its course resembles multinominal graph 6 degrees with three extremes in 1, 4 and 6th hour. 2. Photomorphometric parameters allow for determining the status of the nucleus of hepatocyte in the course of autolysis. 3. The fourth hour of autolysis is a characteristic point after which no response of nucleus to deteriorating oxygen condition was observed. 4. In postmortem autolysis of the nuclei of hepatocytes pyknosis is found to appear, there is no phenomenon of karyorrhexis. 5. After a 7-hour-long experiment, Feulgen's diffusional reaction starts in cytoplasm. 6. A group of tetraploid nuclei is being isolated in the course of the autolysis of hepatocytes. 7. In the course of experiment there appeared a small quantitative loss of nuclear DNA.

摘要

对10只兔肝脏的肝细胞进行了研究。1岁的动物通过枕下区域打击处死。在无菌条件下,将每个肝脏分成9份。其中一份立即检查(零时间),其余部分置于用林格氏液浸湿的纱布衬里的无菌容器中,营造100%湿度的环境。在37摄氏度下进行8小时的自溶,每隔一小时进行一次研究。用于研究的细胞通过制备源自肝段新鲜横截面的接触涂片获得。涂片在40%甲醛蒸汽中于21摄氏度固定2小时,随后在96%乙醇中固定0.5小时。按照常规方法,使用美国菲舍尔公司的碱性品红NI 42510进行福尔根反应。细胞核的形态光度测量借助卡尔·蔡司公司(耶拿,1982年)的“Morphoquant”计算机设备进行。对细胞核进行了以下测量:A. 形态测量——表面积、周长、凸起表面积、直径比、未染色表面积;B. 光度测量——平均消光、周边消光、总消光。研究结果用95张图表记录。根据测量结果确定,在7小时的自溶过程中,肝细胞的细胞核经历动态变化,绘制的函数不呈线性。观察期间,周长和表面积类似于有三个极值的多项式(图12、21、39);同样,平均消光和周边消光也是如此(与表面积和周长相比,其剧烈变化引人关注)(图57、66)。实验时,核群体的分化朝着均匀化方向变化,在或多或少的程度上涉及所有研究特征(比较直方图数据)。四倍体核的孤立群体的特征是占据另一个位置。它由空间排列中的3个变量定义(图76、80)。相关性分析确定了所研究变量之间的动态联系,这些联系似乎特定于给定的实验时间(表1、2)。基于所进行的研究,得出了7条最终结论:1. 肝细胞细胞核的自溶是非线性现象;(其过程类似于6次多项式曲线,在第1、4和6小时有三个极值。)2. 光形态测量参数可用于确定自溶过程中肝细胞细胞核的状态。3. 自溶的第4小时是一个特征点,此后未观察到细胞核对缺氧状况的反应。4. 在肝细胞细胞核的死后自溶中,发现有核固缩现象,没有核碎裂现象。5. 经过7小时的实验,福尔根扩散反应在细胞质中开始。6. 在肝细胞自溶过程中,四倍体核的一组被分离出来。7. 在实验过程中,核DNA出现少量定量损失。

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