METHYLENE DIANILINE TOXICITY TO BILIARY EPITHELIAL CELLS

亚甲基二苯胺(4，4‘-二氨基二苯基甲烷，DAPM)是一种化合物 广泛用于生产环氧树脂和聚氨基甲酸乙酯 目前用于制造主动脉内气球和 人工透析机的管路。DAPM产生选择性毒性 对大鼠胆管上皮细胞(BEC)的作用机制不明。 以前对小剂量DAPM的研究表明，BEC会发生变化 线粒体超微结构与血糖异常升高 胆汁。因为BEC负责从胆汁中吸收葡萄糖，所以我 假设DAPM损伤表面葡萄糖受体和/或改变 高能磷酸盐代谢调节ATP依赖的Na+- 依赖葡萄糖吸收。AIM 1将开发一种体外系统来 原代培养大鼠脐静脉内皮细胞。BEC将被分离和培养在 微载体珠子汇合。然后将珠子灌入1毫升 用于模拟活体条件的柱。这个灌流系统将 提供1)对特定DAPM代谢物影响的控制，2) 我们目标人群的特殊性，BEC，以及3)未来的模式 评估运输机制的研究。目标2和目标3将使用这一点 评估DAPM及其代谢产物对人体健康影响的系统 放射性糖的吸收依赖于葡萄糖受体，并取决于 线粒体损伤，可对葡萄糖产生后续影响 吸收。拟议的目标将使我们进一步了解 DAPM致伤机制及相关专业知识 葡萄糖在BEC中的转运过程。

Methylene dianiline (4,4'-diaminodiphenylmethane, DAPM) is a compound widely used in the production of epoxy resins and polyurethanes which are currently used to make items such as intra-aortic balloons and tubing of artificial dialysis machines. DAPM produces selective toxicity to biliary epithelial cells (BEC) of rats by an unknown mechanism. Previous studies of low dose DAPM have shown alterations in BEC mitochondrial ultrastructure and abnormally high levels of glucose in bile. Because BECs are responsible for glucose absorption from bile, I hypothesize that DAPM damages surface glucose receptors and/or alters high energy phosphate metabolism which regulates ATP-dependent, Na+- dependent glucose absorption. Aim 1 will develop an in vitro system to study primary rat BECs. BECs will be isolated and cultured on microcarrier beads to confluency. Beads will then be perfused in 1 ml columns to simulate in vivo conditions. This perfusion system will provide 1) control over the effects Of specific DAPM metabolites, 2) specificity for our target population, BECs, and 3) a model for future studies to assess transport mechanisms. Aims 2 and 3 will use this system to assess the effects of DAPM and its metabolites on the absorption of radioactive sugars dependent on glucose receptors, and on mitochondrial injury which can have subsequent effects on glucose absorption. The proposed aims will provide further insight into the mechanisms of injury induced by DAPM, and specific knowledge regarding glucose transport processes in BEC.