Cyclosporine A nephrotoxicity studied by the combined application of kidney cell lines, hepatocytes, and endothelial-platelet cocultures.

Molecular toxicology Pub Date : 1987-09-01
A Vickers, J Guertler, C Spaans, C Tapparelli, P Donatsch, V Fischer
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Abstract

Established renal epithelial cell lines of human, pig, and dog origin (293, LLC-PK1, MDCK) were examined in terms of nephrotoxicity and ability to biotransform cyclosporine A (CsA). All three cell lines exhibited a comparable concentration dependent cytotoxicity to CsA treatment. Alterations in cell function included a decreased transport of lysine, an inhibition of growth, and an activation of lysosomal and mitochondrial activity as indicated by the increased uptake of neutral red (NR) and increased reduction of the tetrazolium dye MTT at 1-6 microM CsA. Increased leakage of lactic dehydrogenase and activities of gamma-glutamyl transpeptidase (GGT) and N-acetyl-beta-D-hexosaminidase were observed at 48 h and 12 microM CsA. A discrimination between CsA and the less nephrotoxic cyclosporine-(CsH) was shown for DNA synthesis and NR uptake. The contribution of extrarenal parameters on kidney cell function was studied by the addition of medium from hepatocytes exposed to CsA to the kidney cell lines. A more potent inhibition of DNA synthesis and enhanced reduction of MTT resulted than by addition of equimolar CsA directly to the kidney cells. These data indicate that hepatocyte constituents present in the medium due to CsA treatment affect kidney cell function; additionally, the presence of CsA metabolites may contribute to the CsA-induced nephrotoxicity. The vascular nephrotoxicity induced by CsA, an increased deposition of platelets in the renal arterioles, was mimicked by cocultures of endothelial cell monolayers and platelets. CsA increased the aggregability and adherence of platelets to the endothelial cell monolayers, whereas CsH had no effect.

肾细胞系、肝细胞和内皮-血小板共培养联合应用环孢素A肾毒性研究。
研究了人、猪和狗的肾上皮细胞系(293、LLC-PK1、MDCK)的肾毒性和对环孢素A (CsA)的生物转化能力。所有三种细胞系对CsA处理均表现出相当的浓度依赖性细胞毒性。细胞功能的改变包括赖氨酸运输的减少,生长的抑制,溶酶体和线粒体活性的激活,这表明在1-6微米CsA下中性红(NR)的摄取增加和四氮唑染料MTT的减少增加。乳酸脱氢酶渗漏增加,γ -谷氨酰转肽酶(GGT)和n -乙酰- β - d -己糖氨酸酶活性增加。CsA和肾毒性较小的环孢素-(CsH)在DNA合成和NR摄取方面存在区别。通过将暴露于CsA的肝细胞培养基添加到肾细胞系中,研究了肾外参数对肾细胞功能的贡献。与直接向肾细胞中添加等摩尔CsA相比,其对DNA合成的抑制作用更强,MTT的减少也更明显。这些数据表明,由于CsA处理而存在于培养基中的肝细胞成分影响肾细胞功能;此外,CsA代谢物的存在可能有助于CsA引起的肾毒性。内皮细胞单层和血小板共培养可以模拟CsA诱导的血管肾毒性,即肾小动脉血小板沉积增加。CsA增加了血小板对内皮细胞单层的聚集性和粘附性,而CsH没有影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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