Dantas Rodrigo Tavares, Sampaio Tiago Lima, Lima Dânya Bandeira, Menezes Ramon Róseo Paula Pessoa Bezerra de, Canuto Jader Almeida, Toyama Marcos Hikari, Evangelista Janaína Serra Azul Monteiro, Martins Alice Maria Costa
Departamento de Fisiologia e Farmacologia, Universidade Federal do Ceará, Fortaleza, Ceará, Brazil.
Departmento de Análises Clínicas e Toxicológicas, Universidade Federal do Ceará, Fortaleza, Ceará, Brazil.
Toxicon. 2018 Sep 1;151:24-28. doi: 10.1016/j.toxicon.2018.06.074. Epub 2018 Jun 15.
Acute kidney injury (AKI) is one of the most important complications of bothropic poisoning and its early identification remains challenging. The nephrotoxicity of Bothrops insularis venom (BinsV) was previously described by our research group. In this study, we continued to evaluate the effect of BinsV on kidney function in mice and LLC-MK2 proximal tubule cells, evaluating KIM-1 protein as an early AKI biomarker. Male Swiss mice were inoculated with BinsV intramuscularly and observed for 24 h in a metabolic cage model. Urine and blood were collected for biochemical analyses and the kidneys were examined for oxide-reducing balance and submitted to histological analysis. LLC-MK2 cells incubated with BinsV were assessed for cell viability and cell death mechanism by flow cytometry. Histological analysis of the kidneys indicated AKI and the oxide-reducing analyses demonstrated a decreasing in reduced glutathione (GSH) levels and an increasing on Malondialdehyde (MDA) levels. BinsV was cytotoxic to LLC-MK2 and the cytometry analyses suggested necrosis. Within 24 h after the envenomation, urinary creatinine did not increase, but the urinary levels of KIM-1 increased. In conclusion, we found AKI evidence in the kidney tissue and the increase in the KIM-1 levels suggest it can be used as an early AKI biomarker.
急性肾损伤（AKI）是具窍蝮蛇中毒最重要的并发症之一，其早期识别仍然具有挑战性。我们的研究小组之前曾描述过海岛具窍蝮蛇毒（BinsV）的肾毒性。在本研究中，我们继续评估BinsV对小鼠肾功能和LLC-MK2近端小管细胞的影响，将KIM-1蛋白作为急性肾损伤的早期生物标志物进行评估。雄性瑞士小鼠肌肉注射BinsV，并在代谢笼模型中观察24小时。收集尿液和血液进行生化分析，并检查肾脏的氧化还原平衡，然后进行组织学分析。通过流式细胞术评估与BinsV孵育的LLC-MK2细胞的细胞活力和细胞死亡机制。肾脏的组织学分析表明存在急性肾损伤，氧化还原分析显示还原型谷胱甘肽（GSH）水平降低，丙二醛（MDA）水平升高。BinsV对LLC-MK2具有细胞毒性，流式细胞术分析提示细胞坏死。在中毒后24小时内，尿肌酐没有增加，但尿KIM-1水平升高。总之，我们在肾组织中发现了急性肾损伤的证据，KIM-1水平的升高表明它可作为急性肾损伤的早期生物标志物。
