Massicot F, Martin C, Dutertre-Catella H, Ellouk-Achard S, Pham-Huy C, Thevenin M, Rucay P, Warnet J M, Claude J R
Laboratoire de Toxícologie (EA 207), Université René Descartes (Paris V), France.
Arch Toxicol. 1997;71(8):529-31. doi: 10.1007/s002040050423.
FK506 and cyclosporin A (CsA) are two potent immunosupressants with similar toxicity profile. Nephrotoxicity is the main adverse effect of both compounds. The aim of this study is to compare the in vitro nephrotoxic effects on renal epithelial cell line LLC-PK1 by measuring cell viability and energy status as evaluated by concentrations of ATP and ATP metabolites. Cell viability (expressed as IC50 was assessed via thiazolyl blue (MTT) assay after incubation for 4-24 h with FK506 or CsA. ATP and its metabolites were determined by HPLC after 4 and 6 h incubation with FK506 or CsA alone at the respective IC50. Both FK506 and CsA decreased cell viability to similar extents, in a dose- and time-dependent manner. After 4 h incubation, both drugs decreased ATP levels (-25%) and increased uric acid levels. However, the latter percentage increase was twofold higher with CsA (18%) than with FK506 (9%). The energy charge, calculated according to levels of adenine nucleotides, was decreased by 10% in FK506-treated cells and by 27% in CsA-treated cells. At the end of 6-h incubation, FK506-treated cells maintained ATP levels coupled with energy charge at near control levels whereas the levels were 32% lower in CsA treated cells. Compared to the 4 h-incubation, the increase in uric acid was similar for FK506 but was doubled with CsA. The decrease in cell integrity and ATP depletion induced by CsA in LLC-PK1 cells was only transiently observed with FK506. By preserving energy status, FK506 leads to fewer metabolic disturbances than CsA in the renal epithelial cell line LLC-PK1, demonstrating a minor potential nephrotoxicity.
FK506和环孢素A（CsA）是两种毒性特征相似的强效免疫抑制剂。肾毒性是这两种化合物的主要不良反应。本研究的目的是通过测量细胞活力和能量状态（以ATP和ATP代谢物浓度评估）来比较它们对肾上皮细胞系LLC-PK1的体外肾毒性作用。在用FK506或CsA孵育4-24小时后，通过噻唑蓝（MTT）法评估细胞活力（以IC50表示）。在分别以各自的IC50与FK506或CsA单独孵育4小时和6小时后，通过高效液相色谱法测定ATP及其代谢物。FK506和CsA均以剂量和时间依赖性方式使细胞活力降低到相似程度。孵育4小时后，两种药物均降低了ATP水平（-25%）并升高了尿酸水平。然而，CsA使尿酸水平升高的百分比（18%）是FK506（9%）的两倍。根据腺嘌呤核苷酸水平计算的能量电荷，在FK506处理的细胞中降低了10%，在CsA处理的细胞中降低了27%。在6小时孵育结束时，FK506处理的细胞维持ATP水平以及能量电荷接近对照水平，而CsA处理的细胞中这些水平低32%。与4小时孵育相比，FK506处理的细胞尿酸升高情况相似，但CsA处理的细胞尿酸升高情况翻倍。在LLC-PK1细胞中，CsA诱导的细胞完整性降低和ATP耗竭在FK506处理时仅短暂出现。通过维持能量状态，FK506在肾上皮细胞系LLC-PK1中导致的代谢紊乱比CsA少，表明其潜在肾毒性较小。
