Sorensen E M, Smith N K, Boecker C S, Acosta D
In Vitro. 1984 Oct;20(10):771-9. doi: 10.1007/BF02618293.
Parenchymal hepatocytes from neonatal rats were isolated, cultured about 24 h, exposed to cadmium with or without calcium, and processed for scanning electron microscopy. To assess the severity of cadmium-induced changes, exposed hepatocytes were categorized based upon the extent of morphological damage. Differences in surface blebbing, alterations in microvilli, variations in the degree of swelling, and changes in cell shape were used to categorize the severity of cell damage. A double-blind morphometric analysis (a geometricostatistical processing of two-dimensional data for the collection of three-dimensional information) of cellular changes was conducted for each exposure time and for each concentration of cadmium in the presence or absence of calcium. Significant decreases occurred in the percent relative volume of normal, flattened cells present in cultures exposed for 30 min to 50 or 100 microM cadmium in the absence of calcium. In contrast, the percent relative volume of severely damaged spherical cells was significantly increased after exposure to solutions containing 50 or 100 microM cadmium and lacking calcium. Percent relative volume of intermediate cells (which were slightly swollen and showed changes in microvillar number) was significantly increased following a 30 min exposure to all cadmium concentrations in the absence of calcium. The examination of hepatocytes exposed for 60 min showed that the degree of cadmium-induced cytotoxicity was more severe in the absence of calcium than was the case for the hepatocyte cultures exposed for 30 min: approximately 30% more spherical cells and 30% fewer flattened cells were present if cultures were exposed in the absence of calcium for 60 min compared to those exposed for 30 min. The degree of blebbing was significantly greater at all cadmium concentrations in the absence of calcium. The presence of calcium, therefore, reduced cadmium-induced cytotoxicity in primary cultures of rat hepatocytes subjected to morphometric analysis after scanning electron microscopy.
分离新生大鼠的实质肝细胞，培养约24小时，使其暴露于含或不含钙的镉中，然后进行扫描电子显微镜处理。为评估镉诱导变化的严重程度，根据形态损伤程度对暴露的肝细胞进行分类。表面起泡、微绒毛改变、肿胀程度变化和细胞形状改变被用于对细胞损伤的严重程度进行分类。对每个暴露时间以及存在或不存在钙的情况下每种镉浓度，进行细胞变化的双盲形态计量分析（对二维数据进行几何统计处理以收集三维信息）。在无钙条件下暴露于50或100 microM镉30分钟的培养物中，正常扁平细胞的相对体积百分比显著下降。相比之下，暴露于含50或100 microM镉且无钙的溶液后，严重受损的球形细胞的相对体积百分比显著增加。在无钙条件下暴露于所有镉浓度30分钟后，中间细胞（略有肿胀且微绒毛数量有变化）的相对体积百分比显著增加。对暴露60分钟的肝细胞进行检查发现，无钙时镉诱导的细胞毒性程度比暴露30分钟的肝细胞培养物更严重：与暴露30分钟的培养物相比，如果在无钙条件下暴露60分钟，球形细胞大约多30%，扁平细胞少30%。在无钙条件下，所有镉浓度下的起泡程度都显著更高。因此，在扫描电子显微镜后进行形态计量分析的大鼠肝细胞原代培养中，钙的存在降低了镉诱导的细胞毒性。
