Severin María J, Trebucobich Mara S, Buszniez Patricia, Brandoni Anabel, Torres Adriana M
Área Farmacología , Facultad de Ciencias Bioquímicas y Farmacéuticas. Universidad Nacional de Rosario. CONICET , Argentina . Email:
Toxicol Res (Camb). 2016 Jan 7;5(2):530-538. doi: 10.1039/c5tx00436e. eCollection 2016 Mar 1.
Methotrexate (MTX) belongs to a group of medicines known as antimetabolites. It is commonly used in the treatment of malignant diseases and is prescribed in autoimmune and chronic inflammatory disorders. Along with its effective therapeutic power, MTX has adverse effects on several organs, including the kidney. The organic anion transporter 5 (Oat5) is exclusively localized in the renal apical membrane. Oat5 urinary excretion was proposed as an early biomarker in ischemic and nephrotoxic-induced kidney injury and in renal damage due to vascular calcification in preclinical models. The aim of this study was to evaluate Oat5 renal expression and urinary excretion in rats 48 h after the exposure to different doses of MTX, in comparison with traditional markers of renal injury, such as creatinine and urea plasma levels, protein urinary levels, urinary alkaline phosphatase (AP) activity, fractional excretion of water (FE) and renal histology. Male Wistar rats were treated with a single intraperitoneal injection of MTX at different dosages: 40-80-120-180-360 mg per kg b.w. (M40, M80, M120, M180, M360, = 4, respectively) and experiments were carried out 48 h after MTX administration. Oat5 renal expression was evaluated by western blotting and immunohistochemistry. Traditional parameters were only modified at the higher MTX dose (M360). Conversely, Oat5 urinary excretion was elevated at the middle dose of 80 mg per kg b.w. Oat5 renal expression was modified at the highest dose as well, both in homogenates and in apical membranes. These results suggest that Oat5 urinary excretion might serve as an early biomarker of MTX-induced kidney injury.
甲氨蝶呤（MTX）属于一类被称为抗代谢物的药物。它常用于治疗恶性疾病，也被用于自身免疫性疾病和慢性炎症性疾病的处方治疗。除了其有效的治疗作用外，MTX对包括肾脏在内的多个器官有不良反应。有机阴离子转运体5（Oat5）仅定位于肾顶端膜。在临床前模型中，Oat5尿排泄被提议作为缺血性和肾毒性诱导的肾损伤以及血管钙化导致的肾损伤的早期生物标志物。本研究的目的是评估大鼠在暴露于不同剂量的MTX 48小时后Oat5的肾表达和尿排泄情况，并与肾损伤的传统标志物进行比较，如肌酐和尿素血浆水平、尿蛋白水平、尿碱性磷酸酶（AP）活性、水的分数排泄（FE）和肾脏组织学。雄性Wistar大鼠通过腹腔注射不同剂量的MTX进行治疗：每千克体重40 - 80 - 120 - 180 - 360毫克（分别为M40、M80、M120、M180、M360，每组n = 4），在MTX给药后48小时进行实验。通过蛋白质印迹法和免疫组织化学评估Oat5的肾表达。传统参数仅在较高的MTX剂量（M360）下发生改变。相反，每千克体重80毫克的中等剂量时Oat5尿排泄升高。在最高剂量时，匀浆和顶端膜中的Oat5肾表达也发生了改变。这些结果表明，Oat5尿排泄可能作为MTX诱导的肾损伤的早期生物标志物。
