Silva Adriana Carolina Rodrigues Almeida, de Almeida Breno Fernando Martins, Soeiro Carolina Soares, Ferreira Wagner Luis, de Lima Valéria Marçal Félix, Ciarlini Paulo César
Department of Clinics, Surgery, and Animal Reproduction, College of Veterinary Medicine of Araçatuba, São Paulo State University, 793 Clóvis Pestana Street, 16050-680, Araçatuba, São Paulo, Brazil.
Can J Vet Res. 2013 Apr;77(2):136-41.
Oxidative stress is a key component in the immunosuppression of chronic kidney disease (CKD), and neutrophil function may be impaired by oxidative stress. To test the hypothesis that in uremic dogs with CKD, oxidative stress is increased and neutrophils become less viable and functional, 18 adult dogs with CKD were compared with 15 healthy adult dogs. Blood count and urinalysis were done, and the serum biochemical profile and plasma lipid peroxidation (measurement of thiobarbituric acid reactive substances) were determined with the use of commercial reagents. Plasma total antioxidant capacity (TAC) was measured with a spectrophotometer and commercial reagents, superoxide production with a hydroethidine probe, and the viability and apoptosis of neutrophils with capillary flow cytometry and the annexin V-PE system. The plasma concentrations of cholesterol (P = 0.0415), creatinine (P < 0.0001), and urea (P < 0.0001) were significantly greater in the uremic dogs than in the control dogs. The hematocrit (P = 0.0004), urine specific gravity (P = 0.015), and plasma lipid peroxidation (P < 0.0001) were significantly lower in the dogs that were in late stages of CKD than in the control group. Compared with those isolated from the control group, neutrophils isolated from the CKD group showed a higher rate of spontaneous (0.10 ± 0.05 versus 0.49 ± 0.09; P = 0.0033; median ± standard error of mean) and camptothecin-induced (18.53 ± 4.06 versus 44.67 ± 4.85; P = 0.0066) apoptosis and lower levels of superoxide production in the presence (1278.8 ± 372.8 versus 75.65 ± 86.6; P = 0.0022) and absence (135.29 ± 51.74 versus 41.29 ± 8.38; P = 0.0138) of phorbol-12-myristate-13-acetate stimulation. Thus, oxidative stress and acceleration of apoptosis occurs in dogs with CKD, the apoptosis diminishing the number of viable neutrophils and neutrophil superoxide production.
氧化应激是慢性肾脏病（CKD）免疫抑制的关键组成部分，中性粒细胞功能可能会受到氧化应激的损害。为了验证在患有CKD的尿毒症犬中氧化应激增加且中性粒细胞的活力和功能下降这一假设，将18只患有CKD的成年犬与15只健康成年犬进行了比较。进行了血细胞计数和尿液分析，并使用商业试剂测定了血清生化指标和血浆脂质过氧化（硫代巴比妥酸反应性物质的测量）。用分光光度计和商业试剂测量血浆总抗氧化能力（TAC），用氢乙锭探针测量超氧化物生成，并用毛细管流式细胞术和膜联蛋白V-PE系统测量中性粒细胞的活力和凋亡。尿毒症犬的血浆胆固醇浓度（P = 0.0415）、肌酐浓度（P < 0.0001）和尿素浓度（P < 0.0001）显著高于对照犬。处于CKD晚期的犬的血细胞比容（P = 0.0004）、尿比重（P = 0.015）和血浆脂质过氧化（P < 0.0001）显著低于对照组。与从对照组分离的中性粒细胞相比，从CKD组分离的中性粒细胞显示出自发性凋亡率（0.10±0.05对0.49±0.09；P = 0.0033；中位数±平均标准误差）和喜树碱诱导的凋亡率（18.53±4.06对44.67±4.85；P = 0.0066）更高，并且在存在（1278.8±372.8对75.65±86.6；P = 0.0022）和不存在（135.29±51.74对41.29±8.38；P = 0.0138）佛波醇-12-肉豆蔻酸酯-13-乙酸酯刺激的情况下超氧化物生成水平更低。因此，患有CKD的犬会发生氧化应激和凋亡加速，凋亡会减少存活中性粒细胞的数量以及中性粒细胞超氧化物的生成。
