Makhloufi Camélia, Crescence Lydie, Darbousset Roxane, McKay Nathalie, Massy Ziad A, Dubois Christophe, Panicot-Dubois Laurence, Burtey Stéphane, Poitevin Stéphane
Aix Marseille Univ, INSERM 1263, INRAE, C2VN, Marseille, France.
Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States.
TH Open. 2020 Apr 16;4(2):e66-e76. doi: 10.1055/s-0040-1705138. eCollection 2020 Apr.
The coexistence of bleeding and thrombosis in patients with chronic kidney disease (CKD) is frequent and poorly understood. Mouse models are essential to understand complications of CKD and to develop new therapeutic approaches improving the health of patients. We evaluated the hemostasis in two models of renal insufficiency: adenine-diet and 5/6th nephrectomy (5/6Nx). Compared with 5/6Nx mice, mice fed with 0.25% adenine had more severe renal insufficiency and so higher levels of prothrombotic uremic toxins like indoxyl sulfate. More severe renal inflammation and fibrosis were observed in the adenine group, as demonstrated by histological and reverse transcription quantitative polymerase chain reaction experiments. Liver fibrinogen γ chain expression and level of plasma fibrinogen were increased only in adenine mice. In both CKD mouse models, tissue factor (TF) expression was increased in kidney and aorta extracts. Immunochemistry analysis of kidney sections showed that TF is localized in the vascular walls. Thrombin-antithrombin complexes were significantly increased in plasma from both adenine and 5/6Nx mice. Tail bleeding time increased significantly only in adenine mice, whereas platelet count was not significant altered. Finally, results obtained by intravital microscopy after laser-induced endothelial injury showed impaired platelet function in adenine mice and an increase in fibrin generation in 5/6Nx mice. To summarize, adenine diet causes a more severe renal insufficiency compared with 5/6Nx. The TF upregulation and the hypercoagulable state were observed in both CKD models. Bleeding tendency was observed only in the adenine model of CKD that recapitulates the whole spectrum of hemostasis abnormalities observed in advanced human CKD.
慢性肾脏病(CKD)患者中出血与血栓形成并存的情况很常见,且人们对此了解甚少。小鼠模型对于理解CKD的并发症以及开发改善患者健康的新治疗方法至关重要。我们评估了两种肾功能不全模型中的止血情况:腺嘌呤饮食模型和5/6肾切除(5/6Nx)模型。与5/6Nx小鼠相比,喂食0.25%腺嘌呤的小鼠肾功能不全更严重,因此促血栓形成的尿毒症毒素如硫酸吲哚酚水平更高。通过组织学和逆转录定量聚合酶链反应实验表明,腺嘌呤组观察到更严重的肾脏炎症和纤维化。仅在腺嘌呤小鼠中,肝脏纤维蛋白原γ链表达和血浆纤维蛋白原水平升高。在两种CKD小鼠模型中,肾脏和主动脉提取物中的组织因子(TF)表达均增加。肾脏切片的免疫化学分析表明,TF定位于血管壁。腺嘌呤和5/6Nx小鼠血浆中的凝血酶 - 抗凝血酶复合物均显著增加。仅腺嘌呤小鼠的尾部出血时间显著延长,而血小板计数无显著变化。最后,激光诱导内皮损伤后活体显微镜检查结果显示,腺嘌呤小鼠的血小板功能受损,5/6Nx小鼠的纤维蛋白生成增加。总之,与5/6Nx相比,腺嘌呤饮食导致更严重的肾功能不全。在两种CKD模型中均观察到TF上调和高凝状态。仅在CKD的腺嘌呤模型中观察到出血倾向,该模型概括了晚期人类CKD中观察到的整个止血异常谱。