Valentovic M A, Ball J G, Anestis D K, Rankin G O
Department of Pharmacology, Marshall University School of Medicine, 1542 Spring Valley Drive, Huntington, WV 25755-9310, USA.
Toxicol In Vitro. 1995 Feb;9(1):75-81. doi: 10.1016/0887-2333(94)00188-z.
Acute exposure to certain dichloroaniline (DCA) isomers results in renal and hepatic toxicity in vivo. In the present study we examined whether dichloroaniline structural isomers were cytotoxic to liver and kidney slices in vitro and compared the toxicities of the different structural isomers. These studies were necessary in order to validate the use of an in vitro slice system for examination of the cellular mechanisms for toxicity. Renal cortical and hepatic slices were incubated for 90 min with 2,3-DCA, 2,4-DCA, 2,5-DCA, 2,6-DCA, 3,4-DCA or 3,5-DCA at a final concentration of 0-1 mm. Pyruvate-directed gluconeogenesis was measured following an additional 30-min incubation with 10 mM pyruvate. Cytotoxicity was also determined by measurement of lactate dehydrogenase (LDH) release 120 min after the addition of dichloroaniline isomers at a final concentration of 0, 0.5, 1 or 2 mM. Gluconeogenesis in renal cortical slices was inhibited by all of the isomers beginning at a concentration of 0.5 mM. Renal slice LDH leakage was elevated above control levels by 1-2 mM 3,4-DCA or 3,5-DCA. A final concentration of 2 mM was needed for 2,3-DCA, 2,4-DCA, 2,5-DCA or 2,6-DCA in order to detect a significant (P < 0.05) increase in renal slice LDH leakage. Hepatic slices incubated with 0.5-2 mM 2,3-DCA or 2 mM 2,5-DCA exhibited diminished pyruvate-directed gluconeogenesis. After exposure to 2,4-DCA, 2,6-DCA, 3,4-DCA or 3,5-DCA, pyruvate-directed gluconeogenesis was similar to that in the controls. LDH leakage was increased significantly (P < 0.05) above control values by exposure to 2 mM 3,4-DCA or 3,5-DCA. In conclusion, DCA structural isomers were toxic in vitro to liver and kidney slices. These results indicated that the kidney was more sensitive than the liver to DCA isomers, and that the most toxic isomer was 3,5-DCA. These results are similar to those previously observed in vivo.
急性接触某些二氯苯胺（DCA）异构体在体内会导致肾毒性和肝毒性。在本研究中，我们检测了二氯苯胺结构异构体在体外对肝切片和肾切片是否具有细胞毒性，并比较了不同结构异构体的毒性。进行这些研究是为了验证体外切片系统用于检测毒性细胞机制的有效性。将肾皮质切片和肝切片与2,3 - DCA、2,4 - DCA、2,5 - DCA、2,6 - DCA、3,4 - DCA或3,5 - DCA在终浓度为0 - 1 mM的条件下孵育90分钟。在与10 mM丙酮酸额外孵育30分钟后，测定丙酮酸引导的糖异生作用。在加入终浓度为0、0.5、1或2 mM的二氯苯胺异构体120分钟后，通过测量乳酸脱氢酶（LDH）释放量来确定细胞毒性。所有异构体从0.5 mM浓度开始均抑制肾皮质切片中的糖异生作用。1 - 2 mM的3,4 - DCA或3,5 - DCA使肾切片LDH泄漏量升高至高于对照水平。2,3 - DCA、2,4 - DCA、2,5 - DCA或2,6 - DCA需要终浓度为2 mM才能检测到肾切片LDH泄漏量显著（P < 0.05）增加。用0.5 - 2 mM的2,3 - DCA或2 mM的2,5 - DCA孵育肝切片后，丙酮酸引导的糖异生作用减弱。暴露于2,4 - DCA、2,6 - DCA、3,4 - DCA或3,5 - DCA后，丙酮酸引导的糖异生作用与对照相似。暴露于2 mM的3,4 - DCA或3,5 - DCA使LDH泄漏量显著（P < 0.05）高于对照值。总之，DCA结构异构体在体外对肝切片和肾切片有毒性。这些结果表明，肾脏对DCA异构体比肝脏更敏感，毒性最大的异构体是3,5 - DCA。这些结果与先前在体内观察到的结果相似。
