Development of new methods for the evaluation and protection of drug induced nephrotoxicity using kidney epithelial cell line (LLC-PK_1)

肾脏是排泄外源性物质的主要器官，容易受到药物引起的肾毒性的影响。本研究利用体外培养的人肾上皮细胞株LLC-PK_1，试图建立一种新的评价药物肾毒性和保护药物肾毒性的方法。本研究采用氨基糖苷类抗生素、环孢菌素（免疫抑制剂）和顺铂（抗肿瘤剂）。这些药物在临床上被广泛使用，但它们的使用有时会因急性肾毒性而复杂化。在LLC-PK_1细胞中测试了氨基糖苷、环孢菌素和顺铂的作用，发现活细胞的数量、漂浮细胞（死细胞）的数量、形成圆顶的能力和顶端标记酶（氨肽酶、碱性磷酸酶和γ-谷氨酰转移酶）的活性可用作药物诱导的细胞毒性的标记。各种氨基糖苷类抑制LLC-PK细胞顶端酶的效力之间存在良好的关系 关于我们 以及这些药物在体内的肾毒性。此外，氨基糖苷类和环孢素可引起细胞内游离钙浓度升高，这可能是药物性肾毒性的重要决定因素.庆大霉素对根尖酶活性的抑制作用呈剂量依赖性，并与庆大霉素在细胞内的蓄积有关。根据细胞内游离钙离子浓度和活细胞数判断，环孢素的细胞内蓄积与其细胞毒性也有一定的关系.顺铂的细胞毒性是更有效的指数增长的细胞比汇合细胞。谷氨酰胺可减轻顺铂的毒性。结论：培养的肾上皮细胞系（LLC-PKi）可作为评价药物肾毒性的模型系统，并为药物肾毒性的保护提供新的方法。少

The kidney is a major organ for the excretion of xenobiotics, and is susceptible to drug-induced nephrotoxicity. We attempted to establish new methods for the evaluation and protection of nephrotoxicity of drugs by using a cultured kidney epithelial cell line, LLC-PK_1. Aminoglycoside antibiotics, cyclosporin (immunosuppressive agent), and cisplatin (antitumor agent) were employed in this study. These drugs are widely used clinically, but their use is sometimes complicated by acute nephrotoxicity.1. The effect of aminoglycoside, cyclosporin, and cisplatin was tested in LLC-PK_1 cells, and it was found that the number of viable cells, the number of floating cells (dead cells), the ability to form domes, and the activities of apical marker enzymes (aminopeptidase, alkaline phosphatase, and gamma-glutamyltransferase) are useful as markers of drug-induced cytotoxicity. There was a good relationship between the potencies of various aminoglycosides to inhibit apical enzymes in LLC-PK, cells … More and the nephrotoxic potencies of these drugs in vivo. In addition, aminoglycoside and cyclosporin induced the elevation of cytosolic free calcium concentration, which may be a important determinant of drug-induced nephrotoxicity.2. The inhibitory effect of gentamicin on the apical enzyme activities was dosedependent, and was related to its accumulation in the cells. Apparently, intracellular accumulation of cyclosporin was also related to its cytotoxicity, when judged by the level of cytosolic free calcium and the number of viable cells.3. Cytotoxicity of cisplatin was more potent on exponentially growing cells than on confluent cells. Glutathione attenuated the toxicity of cisplatin. In contrast, cyclosporin was toxic on growing and confluent cells, and glutathione had no effect on its cytotoxicity, indicating that biochemical mechanisms of nephrotoxicity are different among different drugs.In conclusion, cultured kidney epithelial cell line (LLC-PKi) should be a useful model system to evaluate the nephrotoxicity of drugs, and to develop a new method to protect the drug-induced nephrotoxicity. Less