Toutain H J, Sarsat J P, Bouant A, Hoet D, Leroy D, Moronvalle-Halley V
Département Sécurité du Médicament, Rhône-Poulenc Rorer SA, Vitry sur Seine, France.
Cell Biol Toxicol. 1996 Dec;12(4-6):289-98. doi: 10.1007/BF00438160.
The dog is the non-rodent species the most often used in preclinical drug safety evaluation. In this study, we established a new system of precision-cut dog renal cortical slices, evaluated their biochemical, functional, and morphological integrity, and determined the effects of cisplatin (cis-diamminedichloroplatinum (II), CDDP), a very potent nephrotoxic antineoplastic agent used to treat a variety of solid tumors, on the viability and histopathology of slices. Precision-cut renal cortical slices were made perpendicular to the cortical-papillary axis. Slices were incubated in DMEM/Ham's F12 culture medium containing 1 g/L glucose, 2 mmol/L glutamine, and 2 mmol/L heptanoic acid at 37 degrees C in an atmosphere of 5% CO2-70% O2-25% N2 in dynamic organ culture. Our results showed that slices maintained ATP and GSH content, protein synthesis, Na(+)-dependent uptake of glucose inhibited by phlorizin, PAH (p-aminohippuric acid) uptake inhibited by probenecid, and TEA (tetraethylammonium) uptake inhibited by mepiperphenidol for at least 6 h of culture, and morphological integrity up to 24 h. After 6 h of exposure, CDDP induced vacuolation and cell necrosis in the epithelial tubular cells of slices with a concentration-related increase in extension but not in severity. The development of the lesions started in the proximal tubules and extended to the distal tubules. The location and the extension of the lesions confirmed the observations in dog kidneys after in vivo treatment with CDDP by the intravenous route. The concentration-related decrease in slice viability after 6 h exposure to CDDP was in keeping with the extension of the histopathological lesions in the renal parenchyma. The slice viability was unaffected up to 0.63 mmol/L CDDP. At 1.25 and 2.5 mmol/L CDDP, slice viability fell by 35% and 75%, respectively. These results suggest that precision-cut dog renal cortical slices in culture may be suitable for addressing the specific nephrotoxicity issues encountered in this species.
狗是临床前药物安全性评价中最常使用的非啮齿类动物。在本研究中，我们建立了一种新的精确切割狗肾皮质切片系统，评估了它们的生化、功能和形态完整性，并确定了顺铂（顺二氨二氯铂（II），CDDP），一种用于治疗多种实体瘤的极具肾毒性的抗肿瘤药物，对切片活力和组织病理学的影响。精确切割的肾皮质切片垂直于皮质 - 乳头轴制作。切片在含有1 g/L葡萄糖、2 mmol/L谷氨酰胺和2 mmol/L庚酸的DMEM/Ham's F12培养基中，于37℃、5% CO2 - 70% O2 - 25% N2的气氛中进行动态器官培养。我们的结果表明，切片在培养至少6小时内维持了ATP和谷胱甘肽含量、蛋白质合成、根皮苷抑制的Na(+)依赖性葡萄糖摄取、丙磺舒抑制的对氨基马尿酸（PAH）摄取以及美哌隆抑制的四乙铵（TEA）摄取，并且在长达24小时内保持形态完整性。暴露6小时后，CDDP诱导切片上皮肾小管细胞出现空泡化和细胞坏死，其扩展呈浓度依赖性增加，但严重程度无增加。病变从近端小管开始发展并延伸至远端小管。病变的位置和扩展证实了静脉注射CDDP后在狗肾脏中的体内观察结果。暴露于CDDP 6小时后，切片活力的浓度依赖性下降与肾实质中组织病理学病变的扩展一致。在0.63 mmol/L CDDP时，切片活力不受影响。在1.25和2.5 mmol/L CDDP时，切片活力分别下降了35%和75%。这些结果表明，培养的精确切割狗肾皮质切片可能适用于解决该物种中遇到的特定肾毒性问题。
