Mitchell D B, Acosta D
J Toxicol Environ Health. 1981 Jan;7(1):83-92. doi: 10.1080/15287398109529960.
Primary cultures of hepatocytes from postnatal Sprague-Dawley rats were grown in arginine-deficient, ornithine-supplemented medium to inhibit fibroblastic overgrowth and to selectively isolate relatively pure cultures of parenchymal hepatocytes. This system of primary cultures of rat hepatocytes was utilized to evaluate the cytotoxicity of certain tricyclic antidepressant drugs (TCAs). The compounds tested were chosen to represent two distinct chemical classifications of TCAs: the dibenzazepine derivatives, imipramine (1) and desipramine (D), and the dibenzocycloheptadiene derivatives, amitriptyline (A) and nortriptyline (N). The study also allowed direct comparison of the parent tertiary amines, A and I, and their respective demethylated pharmacologically active metabolites, N and D. The hepatotoxicity of the compounds was determined by measuring leakage of cytoplasmic enzymes, lactate dehydrogenase (LDH) and glutamic-pyruvic transaminase (GPT), into the culture medium and by assessing cell viability by the trypan blue dye exclusion test. LDH leakage was a more sensitive index of early cellular injury in this study. The compounds demonstrated a dose- and time-dependent order of toxicity; their hepatotoxicity potency was ranked as A = N greater than D greater than I.
将出生后的斯普拉格-道利大鼠的肝细胞进行原代培养，培养于缺乏精氨酸但补充鸟氨酸的培养基中，以抑制成纤维细胞过度生长，并选择性地分离出相对纯净的实质肝细胞培养物。利用该大鼠肝细胞原代培养系统评估某些三环类抗抑郁药物（TCA）的细胞毒性。所测试的化合物被选来代表三环类抗抑郁药物的两种不同化学分类：二苯并氮杂卓衍生物，丙咪嗪（1）和地昔帕明（D），以及二苯并环庚二烯衍生物，阿米替林（A）和去甲替林（N）。该研究还允许直接比较母体叔胺，A和I，以及它们各自的去甲基化药理活性代谢物，N和D。通过测量细胞质酶乳酸脱氢酶（LDH）和谷丙转氨酶（GPT）泄漏到培养基中的情况，并通过台盼蓝染料排除试验评估细胞活力，来确定化合物的肝毒性。在本研究中，LDH泄漏是早期细胞损伤的更敏感指标。这些化合物表现出剂量和时间依赖性的毒性顺序；它们的肝毒性效力排名为A = N大于D大于I。
