In vitro response of rat and human kidney lysosomes to aminoglycosides.
In vitro response of rat and human kidney lysosomes to aminoglycosides.
复制标题
大鼠和人肾溶酶体对氨基糖苷类药物的体外反应。
DOI：
10.1016/0006-2952(82)90625-6
复制
发表时间：
1982
影响因子：
5.8
通讯作者：
Reidenberg,MM
中科院分区：
文献类型：
--
作者：
Powell,JH;Reidenberg,MM
为了研究氨基糖苷的肾毒性，从大鼠肾脏和5个因肿瘤切除的人肾脏的健康部分制备了肾皮质溶酶体。肾皮质用1 mM EDTA和0.3M蔗糖匀浆，差速离心分离溶酶体。溶酶体在不同药物浓度的等渗蔗糖溶液中37°孵育1小时。将它们重新沉积，并在上清部分和破碎的颗粒中测量乙酰-β-氨基葡萄糖酶（NAG）的活性。用治疗血浆浓度的四种氨基糖苷进行教育，以剂量相关的方式降低了NAG释放到上清部分的百分比。与精胺和亚精胺孵育也产生这种效果，精胺和庆大霉素是添加剂。在用等渗甘氨酸代替蔗糖后，也观察到溶酶体在临床达到的血浆浓度下的明显稳定。高浓度的氨基糖苷与患者和大鼠肾皮质中积累的氨基糖苷一致，产生溶酶体NAG的剂量依赖性释放，其效价的等级顺序与临床观察到的产生肾毒性的潜力平行。大鼠给予20 mg/kg庆大霉素，每天2次，连续28天，产生对氨基糖苷肾毒性的肾脏抗性。与盐水处理的对照组相比，这些动物制备的溶酶体对庆大霉素（2和4μg/ml）和亚胺的反应降低。人肾皮质溶酶体也表现出氨基糖苷和多胺诱导的NAG释放变化。我们认为溶酶体是氨基糖苷肾毒性的作用位点。我们认为这种溶酶体膜的氨基糖苷稳定性可能导致近端小管溶酶体系统的最终破坏和细胞损伤。
To study aminoglycoside nephrotoxicity, renal cortical lysosomes were prepared from rat kidneys and from healthy portions of five human kidneys removed for tumor. The renal cortex was homogenized in 1 mM EDTA with 0.3M sucrose, and the lysosomes were separated by differential centrifugation. Lysosomes were incubated in isotonic sucrose solution with various drug concentrations for 1 hr at 37°. They were resedimented adn theN-acetyl-β-glucosaminidase (NAG) activity was measured in the supernatant fraction and in the disrupted pellet. Incucation with four aminoglycosides at therapeutic plasma concentrations lowered the percentage of NAG released into the supernatant fraction in a dose-related fashion. Incubation with the polyamines spermine and spermidine also produced this effect, with spermine and gentamicin being additive. This apparent lysosomal stabilization at clinically achieved plasma concentrations was also observed after substituting isotonic glycine for sucrose in the incubation mixture. High concentrations of aminoglycoside consistent with those accumulated in the renal cortex of patients and rats produced a dose-dependent release of lysosomal NAG with a rank order of potency paralleling their clinically observed potential for producing nephrotoxicity. Rats were treated with 20 mg/kg gentamicin twice a day for 28 days producing kidneys resistant to aminoglycoside nephrotoxicity. Lysosomes prepared from these animals compared to saline-treated controls showed decreased response to gentamicin at 2 and 4μg/ml and to apermine. Human renal cortical lysososomes also exhibited aminoglycoside- and polyamine-induced changes in NAG release. We conclude taht the lysosome is a site of action for aminoglycoside nephrotoxicity. We propose that aminoglycoside stabilization of this lysosomal membrane may lead to eventual disruption of the proximal tubular lysosomal system and cell injury.