Sorensen E M
College of Pharmacy, Department of Pharmacology and Toxicology, University of Texas, Austin 78712.
Toxicol Lett. 1989 Jan;45(1):101-10. doi: 10.1016/0378-4274(89)90164-1.
Morphometric analysis was used to quantitate indomethacin-induced morphological changes in primary cultures of neonatal rat hepatocytes by comparison with standard assessments of functional integrity (i.e. enzyme release, urea levels, and dye exclusion). For this procedural validation, indomethacin concentrations were selected to correspond to the therapeutic plasma levels of rheumatoid arthritic or systemic lupus erythematosus patients having liver cell injury secondary to prolonged, high-dose administration of this nonsteroidal antiinflammatory agent. Primary cultures of neonatal rat hepatocytes were exposed for 12 h to 0, 100, 500 or 1000 microM indomethacin and subjected to a double-blind morphometric analysis procedure modified for use on cultured cells. This procedure systematically converted two-dimensional morphologic information into three-dimensional numerical data for statistical analysis. These optical measurements provided an accurate analysis following 4 h of measurements. When the concentration of indomethacin was increased from 0 to 1000 microM, the relative volume percent of Type I cells decreased. Therefore these cells, which were indistinguishable from healthy, untreated control cells, become less numerous as toxicant level increased. In contrast, the relative volume percent values of other progressively more damaged cell types (i.e. Types II, III, and IV cells) increased with elevation of indomethacin levels. Morphometric assessments paralleled functional assessments of indomethacin-induced cytotoxicity (r2: 0.88-0.99). Therefore, the present study validated this morphometric analysis procedure using a rigorous cellular exposure which caused substantial cell injury and subsequent lifting of 23% cells from the substrate. Combined with previous validation studies involving cadmium, erythromycin and benoxaprofen, the present study showed that morphometric analysis is a rapid, accurate method for the measurement of cell injury in cultured parenchymal hepatocytes.
通过与功能完整性的标准评估（即酶释放、尿素水平和染料排斥）进行比较，形态计量分析用于定量吲哚美辛诱导的新生大鼠肝细胞原代培养物中的形态变化。为了进行该程序验证，选择的吲哚美辛浓度对应于类风湿性关节炎或系统性红斑狼疮患者因长期高剂量服用这种非甾体抗炎药而继发肝细胞损伤时的治疗性血浆水平。将新生大鼠肝细胞原代培养物暴露于0、100、500或1000微摩尔的吲哚美辛中12小时，并进行经修改以用于培养细胞的双盲形态计量分析程序。该程序系统地将二维形态学信息转换为三维数值数据以进行统计分析。在测量4小时后，这些光学测量提供了准确的分析。当吲哚美辛浓度从0增加到1000微摩尔时，I型细胞的相对体积百分比降低。因此，这些与健康、未处理的对照细胞无法区分的细胞，随着毒物水平的增加而数量减少。相比之下，其他逐渐受损更严重的细胞类型（即II型、III型和IV型细胞）的相对体积百分比值随着吲哚美辛水平的升高而增加。形态计量评估与吲哚美辛诱导的细胞毒性的功能评估平行（r2：0.88 - 0.99）。因此，本研究使用严格的细胞暴露验证了这种形态计量分析程序，该暴露导致了大量细胞损伤并随后有23%的细胞从底物上脱离。结合先前涉及镉、红霉素和贝诺洛芬的验证研究，本研究表明形态计量分析是一种快速、准确的方法，用于测量培养的实质肝细胞中的细胞损伤。
