Wallace K A, Harbell J W, Accomando N, Triana A, Valone S, Curren R D
Microbiological Associates Inc., 9900 Blackwell Rd, Rockville, MD 20850, USA.
Toxicol In Vitro. 1992 Jul;6(4):367-71. doi: 10.1016/0887-2333(92)90027-o.
Two methodologies used in vitro to estimate cytotoxicity in cell culture systems were compared: these were the neutral red uptake assay (NRU), which is used to measure toxicity caused by an extended (48-hr) exposure to the test material, and the neutral red release assay (NRR), which is used to measure toxicity caused by a short-term (1-min) exposure to the test material. Both methodologies used the normal human epidermal keratinocyte (NHEK)-based NeutralRed Bioassay supplied by Clonetics Corporation (San Diego, CA, USA). 10 materials (paracetamol, acetylsalicylic acid, ferrous sulphate, diazepam, amitriptyline, digoxin, ethylene glycol, methanol, ethanol and isopropanol), which are part of the Multicenter Evaluation of In Vitro Cytotoxicity (MEIC) panel, were tested. NRU(50) values for the 10 compounds covered more than an eight-log range from 0.004 mum (digoxin) to 1.0 x 10(6) mum (methanol). Because of solubility limits, NRR(50) values for diazepam, digoxin, ferrous sulphate and paracetamol could not be determined. NRR(50) values for the remaining six compounds covered approximately a three-log range from 3.2 x 10(3) to 7.1 x 10(6) mum. When compared with documented values for either the human acute oral lethal dose or the human acute lethal blood concentration, the NRU assay was found to be much more useful in predicting human acute toxicity than was the NRR assay.
一种是中性红摄取试验（NRU），用于测量长时间（48小时）接触测试材料所引起的毒性；另一种是中性红释放试验（NRR），用于测量短时间（1分钟）接触测试材料所引起的毒性。两种方法均使用了由美国加利福尼亚州圣地亚哥的Clonetics公司提供的基于正常人表皮角质形成细胞（NHEK）的中性红生物测定法。测试了多中心体外细胞毒性评估（MEIC）小组中的10种材料（对乙酰氨基酚、乙酰水杨酸、硫酸亚铁、地西泮、阿米替林、地高辛、乙二醇、甲醇、乙醇和异丙醇）。这10种化合物的NRU(50)值涵盖了超过8个对数范围，从0.004 μmol（地高辛）到1.0×10⁶ μmol（甲醇）。由于溶解度限制，无法确定地西泮、地高辛、硫酸亚铁和对乙酰氨基酚的NRR(50)值。其余6种化合物的NRR(50)值涵盖了大约3个对数范围，从3.2×10³到7.1×10⁶ μmol。与人类急性口服致死剂量或人类急性致死血药浓度的记录值相比，发现NRU试验在预测人类急性毒性方面比NRR试验更有用。
