目的

建立并比较3 种急性肾脏病（AKD）动物模型，为研究AKD 病理生理机制提供基础研究证据。

方法

选取6～8 周健康雄性C57BL/6J 小鼠90 只，按照建立3 种AKD 模型，各分为模型组和对照组（15 只/组）：叶酸模型组（给予0.5%叶酸250 mg/kg 腹腔注射）、叶酸对照组（等量0.3 M 碳酸氢钠腹腔注射）；单侧输尿管梗阻（UUO）模型组（行单侧输尿管结扎）、UUO 对照组（仅游离输尿管）；单侧缺血再灌注（UIR）模型组（钳夹单侧肾蒂30 min）、UIR 对照组（仅游离单侧肾蒂）。建模后2 d、8 d 和14 d 分别取材，检测血清肌酐、血尿素氮、蛋白尿水平，观测肾组织病理并评估肾损伤及胶原沉积。 Western 印迹检测纤维化相关的波形蛋白和α-平滑肌肌动蛋白表达；RT-qPCR 检测肾组织的肾损伤分子（KIM-1）及炎症因子（TNF-α、IL-1β、IL-8、IFN-γ）mRNA 表达。

结果

叶酸模型组在建模后8 d 时血清肌酐和血尿素氮明显升高（P 均＜0.05），在建模后2 d 时KIM-1 表达高于其他两个模型组。 UUO 模型组在建模后8 d 时KIM-1 表达则高于其他两个模型组（P 均＜0.05）。 3 个模型组的肾小管损伤分数和胶原沉积呈现时间依赖性增多，且均多于各自对照组。 在建模后14 d 时，UUO 模型组肾小管损伤分数、胶原沉积、波形蛋白和α-平滑肌肌动蛋白表达均高于另外两个模型组（P 均＜0.05）。 在建模后8 d 时，3 个模型组的IL-8、IL-1β、TNF-α 和IFN-γ mRNA 表达均高于其对照组（P 均＜0.05），而UUO 模型组的这些炎症因子表达高于另外两个模型组（P＜0.05）。

结论

小鼠AKD 的叶酸模型、UUO 模型和UIR 模型均在建模操作后8 d 建成，其中UUO 模型在疾病进展中的肾纤维化更为严重。

Objective

To establish and compare three animal models of acute kidney disease（AKD） to provide basic research evidence for investigating the pathophysiological mechanisms of AKD.

Methods

Ninety healthy male C57BL/6J mice aged 6-8 weeks were selected， and divided into model groups and control groups （15 mice/group） to establish three AKD models： folic acid model group （given intraperitoneal injection of 0.5% folic acid at 250 mg/kg）， folic acid control group （given intraperitoneal injection of the same amount of 0.3 M sodium bicarbonate）， unilateral ureteral obstruction （UUO） model group （given unilateral ureteral obstruction）， UUO control group （given sham operation）， unilateral ischemia-reperfusion （UIR） model group （given clamping of unilateral renal pedicle for 30 minutes）， and UIR control group （given sham operation）.On day 2， day 8， and day 14 after the modeling， samples were collected to measure serum creatinine， blood urea nitrogen， and proteinuria levels.Renal tissue pathology was observed， and renal injury and collagen deposition were assessed.Western blotting was used to detect the expressions of fibrosis-related proteins such as vimentin and α-smooth muscle actin.RT-qPCR was used to detect renal mRNA expressions of kidney injury molecule-1 （KIM-1） and inflammatory factors including TNF-α， IL-1β， IL-8， and IFN-γ）.

Results

In the folic acid model group， serum creatinine and blood urea nitrogen significantly increased at day 8 after the modeling （both P＜0.05）， while the KIM-1 expression was higher than those in the other two model groups at day 2 after the modelings.At day 8 after the modelings， the expression of KIM-1 in the UUO model group was higher than those in the other two model groups （all P＜0.05）.The tubular injury scores and collagen deposition in the three model groups showed a time-dependent increase， and were all higher than those in their respective control groups.At day 14 after modeling， the UUO model group showed higher tubular injury scores and collagen deposition， as well as higher expressions of vimentin and α-smooth muscle actin than the other two model groups （all P＜0.05）.At day 8 after the modelings， the mRNA expressions of IL-8， IL-1β， TNF-α， and IFN-γ in the three model groups were all higher than those in their respective control groups （all P＜0.05）， while the expressions of these inflammatory factors in the UUO model group were higher than those in the other two model groups （all P＜0.05）.

Conclusion

The folic acid model， UUO model， and UIR model of mice AKD were all established at day8after the modeling operations， among which the renal fibrosis during the disease progression was more severe in the UUO model.