七叶皂苷钠对单侧输尿管梗阻后小鼠肾积水损伤的保护作用

Protective effect of sodium aescinate against hydronephrosis injury in mice after unilateral ureteral obstruction

摘要:

目的 探讨七叶皂苷钠对小鼠单侧输尿管梗阻（unilateral ureteral obstruction，UUO）后肾积水损伤的改善及保护作用。方法 随机挑选24只健康雄性8周龄WTC57BL/6小鼠，将其分为假手术组（Sham组）、模型组（单侧输尿管梗阻组，UUO组）和七叶皂苷钠用药组（UUO+七叶皂苷钠组），每组8只。UUO模型为肾积水研究最常用模型，采取腰部切口并丝线结扎一侧输尿管（结扎单侧输尿管，并切除对侧肾脏，以确保单侧出现肾积水，并排除对侧肾脏正常代偿的影响），构建单侧输尿管梗阻小鼠肾积水模型，对各组进行相应处理后，全自动生化分析仪检测血清肌酐（serum creatinine，Scr）、尿素氮（blood urea nitrogen，BUN）水平；试剂盒检测肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）、超氧化物歧化酶（superoxide dismutase，SOD）、丙二醛（malondialdehyde，MDA）含量；制备组织病理切片并行HE染色，光镜下观察肾组织病理学改变并做出对应的病理损伤评分；TUNEL法检测细胞凋亡水平。结果 与Sham组比较，UUO组Scr（21.49±2.87 vs. 11.21±1.79）、BUN（19.80±2.55 vs. 9.40±1.78）、TNF-α（20.06±1.32 vs. 10.18±0.56），IL-1β（9.65±0.79 vs. 3.46±0.59），MDA（26.01±1.02 vs. 15.12±1.15），细胞凋亡水平（150.02±14.78 vs. 86.82±5.66），肾病理损伤评分（3.00±0.46 vs. 0.00±0.00）水平显著升高（P=0.000），以及显著降低的SOD水平（95.63±3.13 vs. 143.66±5.44，P=0.000）。与UUO组相比，UUO+七叶皂苷钠组Scr（15.94±0.74 vs. 21.49±2.87）、BUN（13.08±0.85 vs. 19.80±2.55），TNF-α（15.01±0.90 vs. 20.06±1.32），IL-1β（5.62±0.65 vs. 9.65±0.79），MDA（19.12±1.08 vs. 26.01±1.02）、细胞凋亡水平（103.97±10.33 vs. 150.02±14.78）、肾病理损伤评分（1.57±0.36 vs. 3.00±0.46）水平显著降低（P=0.000），以及显著增加SOD水平（121.04±3.61 vs. 95.63±3.13，P=0.000）。结论 七叶皂苷钠通过发挥其抗炎、抑制氧化应激反应以及抗渗出以扩张血管张力作用，从而对UUO后肾积水损伤产生保护作用，成为一种潜在的治疗选择。

Abstract:

Objective To investigate the role of sodium aescinate in improving and protecting against hydronephrosis injury in mice after unilateral ureteral obstruction（UUO）.Methods A total of 24 healthy male WTC57BL/6 mice，aged 8 weeks，were divided into sham-operation group（Sham group），model group（UUO group），and sodium aescinate group（UUO+sodium aescinate group），with 8 mice in each group. The UUO model was the most commonly used model for the study of hydronephrosis，and the ureter at one side was ligated with silk thread through a lumbar incision（the ureter at one side was ligated and the kidney at the contralateral side was removed to ensure unilateral hydronephrosis and rule out the influence of normal compensation of the kidney at the contralateral side） to establish a mouse model of hydronephrosis after UUO. After corresponding treatment was given for each group，an automatic biochemical analyzer was used to measure the levels of serum creatinine（Scr） and blood urea nitrogen（BUN）；related kits were used to measure the content of tumor necrosis factor-α（TNF-α），interleukin-1β（IL-1β），superoxide dismutase（SOD），and malondialdehyde（MDA）；histopathological sections were prepared and then HE staining was used to observe renal histopathological changes under a light microscope and determine the corresponding pathological injury score；the TUNEL method was used to measure the level of cell apoptosis.Results Compared with the Sham group，the UUO group had significant increases in the levels of Scr（21.49±2.87 vs. 11.21±1.79），BUN（19.80±2.55 vs. 9.40±1.78），TNF-α（20.06±1.32 vs. 10.18±0.56），IL-1β（9.65±0.79 vs. 3.46±0.59），and MDA（26.01±1.02 vs. 15.12±1.15），the level of cell apoptosis（150.02±14.78 vs. 86.82±5.66），and renal pathological injury score（3.00±0.46 vs. 0.00±0.00）（P=0.000），as well as a significant reduction in the level of SOD（95.63±3.13 vs. 143.66±5.44，P=0.000）. Compared with the UUO group，the UUO+sodium aescinate group had significant reductions in the levels of Scr（15.94±0.74 vs. 21.49±2.87），BUN（13.08±0.85 vs. 19.80±2.55），TNF-α（15.01±0.90 vs. 20.06±1.32），IL-1β（5.62±0.65 vs. 9.65±0.79），and MDA（19.12±1.08 vs. 26.01±1.02），the level of cell apoptosis（103.97±10.33 vs. 150.02±14.78），and renal pathological injury score（1.57±0.36 vs. 3.00±0.46）（P=0.000），as well as a significant increase in the level of SOD（121.04±3.61 vs. 95.63±3.13，P=0.000）.Conclusion Sodium aescinate exerts a protective effect against hydronephrosis injury after UUO by exerting an anti-inflammatory effect，inhibiting oxidative stress response，and showing an anti-exudation effect to increase vascular tension，and therefore，it becomes a potential treatment option.