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2018 年第 8 期 第 13 卷

侧腰部二次手术法建立小鼠慢性肾脏病基础上急性肾损伤模型

Establishment of acute kidney injury mouse model based on chronic kidney disease with a two-step lateral lumbar surgery

作者:高绪霞韩丽媛马立萍王以新

英文作者:

单位:100029首都医科大学附属北京安贞医院全科医疗科(高绪霞、马立萍、王以新),病理科(韩丽媛)

英文单位:

关键词:急性肾损伤;慢性肾脏病;单侧输尿管结扎术;双侧肾脏缺血再灌注手术;动物模型

英文关键词:

  • 摘要:
  • 【摘要】目的    通过侧腰部二次手术法建立小鼠慢性肾脏病基础上急性肾损伤(AKI)的动物模型,为进一步深入研究AKI病理生理机制、探索有效的防治方法构建研究工具。方法    8~12周龄的雄性C57BL/6小鼠3只作为正常对照,16只在麻醉状态下行左侧腰部单侧输尿管结扎术(UUO),48和96 h后各有8只再从双侧腰部行肾脏缺血40 min再灌注24 h手术,记为UUO 3 d+缺血再灌注(I/R)组和UUO 5 d+I/R组。自动生化仪苦味酸法检测血清肌酐水平,左侧肾脏组织石蜡包埋,切成4 μm厚切片,行苏木精-伊红及Masson染色观察,并进行急性小管损伤评分和纤维组织半定量分析。结果    UUO 3 d+I/R组、UUO 5 d+I/R组血清肌酐水平明显高于对照组[(197±47)、(186±26)μmol/L比(54±10)μmol/L],肾脏小管急性损伤评分明显高于对照组,差异均有统计学意义(均P<0.05)。苏木精-伊红染色显示肾脏间质大量炎症细胞浸润、管腔扩张、充斥管型,刷状缘消失、小管上皮细胞空泡变性、坏死脱落等急性小管损伤,并存皮质变薄、小管萎缩、上皮细胞扁平、间质增宽等慢性病变。Masson染色可见UUO 3 d+I/R组、UUO 5 d+I/R组小鼠肾脏间质纤维基质增多。结论    小鼠侧腰部UUO后再行I/R二次手术法可建立慢性肾脏病基础上AKI的模型。

  • Objective    To establish the mouse model of acute kidney injury(AKI) on the basis of chronic kidney disease through a two-step lateral lumbar surgery method. Methods    Male C57BL/6 mice with age of 8-12 weeks old were used in this study. Three mice were normal control. Sixteen mice had unilateral ureteral obstruction(UUO) with left lateral lumbar incision; 8 mice had bilateral renal ischemia(40 min) reperfusion(24 h)(I/R) at 48 h and 96 h after UUO, as UUO 3 d+I/R group and UUO 5 d+I/R group. Serum creatinine was tested by picric acid method. Left renal tissue was separated for hematoxylin eosin(HE) and Masson staining, acute renal tubular injury assessment and fibrous tissue semi-quantitative analysis. Results    Serum creatinine level[(197±47),(186±26)μmol/L vs (54±10)μmol/L] and acute renal tubular injury score in UUO 3 d+I/R group and UUO 5 d+I/R group were significantly higher than those in normal control group(P<0.05). HE staining showed massive inflammatory cell infiltration, tubular dilatation, cast formation, brush border disappearance, tubular epithelial cell vacuolar degeneration and necrosis in renal interstitium, with thin renal cortex, shrinked tubules, flat epithelial cells and broaden interstitial space. Masson staining showed that fiber matrix increased in renal interstitium of mice in UUO 3 d+I/R group and UUO 5 d+I/R group. Conclusion    UUO plus renal ischemia reperfusion is an effective method to establish mouse chronic kidney disease and AKI model.

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