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经膀胱颈部分结扎建立膀胱输尿管返流性肾损害动物模型

王钊颖 张朝霞 金黎明 刘博 沈炼桔 何大维 魏光辉

王钊颖, 张朝霞, 金黎明, 等. 经膀胱颈部分结扎建立膀胱输尿管返流性肾损害动物模型[J]. 四川大学学报(医学版), 2021, 52(5): 825-831. doi: 10.12182/20210960502
引用本文: 王钊颖, 张朝霞, 金黎明, 等. 经膀胱颈部分结扎建立膀胱输尿管返流性肾损害动物模型[J]. 四川大学学报(医学版), 2021, 52(5): 825-831. doi: 10.12182/20210960502
WANG Zhao-ying, ZHANG Zhao-xia, JIN Li-ming, et al. Establishment of an Animal Model of Vesicoureteral Reflux Renal Injury through Partial Bladder Outlet Obstruction[J]. JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION), 2021, 52(5): 825-831. doi: 10.12182/20210960502
Citation: WANG Zhao-ying, ZHANG Zhao-xia, JIN Li-ming, et al. Establishment of an Animal Model of Vesicoureteral Reflux Renal Injury through Partial Bladder Outlet Obstruction[J]. JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION), 2021, 52(5): 825-831. doi: 10.12182/20210960502

经膀胱颈部分结扎建立膀胱输尿管返流性肾损害动物模型

doi: 10.12182/20210960502
基金项目: 国家自然科学基金(No. 81771566)和重庆市科技局技术创新与应用发展(重点项目)(No. cstc2019jscx-tjsbX0003)资助
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    E-mail:u806806@cqmu.edu.cn

Establishment of an Animal Model of Vesicoureteral Reflux Renal Injury through Partial Bladder Outlet Obstruction

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  • 摘要:   目的  建立一种膀胱出口梗阻导致的返流性肾损害动物模型。  方法  取6~8周龄雄性C57BL/6小鼠60只,随机划分为对照组、假手术组和膀胱颈部分结扎(PBOO)组,每组20只,PBOO组小鼠麻醉后开腹分离膀胱颈,用0.3 mm直径的金属棒引导部分结扎膀胱颈,假手术组仅开腹分离膀胱颈,不结扎,对照组不做处理。术后7 d,每组随机挑选12只存活的小鼠,观察膀胱、输尿管、肾盂和肾脏大体变化,行经膀胱逆行性尿路造影,取肾组织进行组织病理学分析,Western blot、免疫荧光和免疫组化分别检测波形蛋白、增殖细胞核抗原(PCNA)、α平滑肌肌动蛋白 (α-SMA)表达水平。  结果  与对照组和假手术组相比,PBOO组小鼠膀胱、输尿管、肾盂明显扩张,膀胱输尿管返流明显,肾脏体积和质量增加(P<0.001),肾实质变薄(P<0.000 1);组织病理学检查可见肾小球萎缩,肾小管扩张,小管间质炎细胞浸润,肾小球基底膜增生,间质纤维化明显;Western blot、免疫荧光和免疫组化分别测得肾组织中波形蛋白、PCNA、α-SMA的表达水平均升高(P<0.000 1)。  结论  小鼠行PBOO术后,膀胱、输尿管、肾脏均发生明显形态改变,并出现返流性肾纤维化样损害,可以作为研究膀胱出口梗阻导致的肾纤维化病理改变机制及治疗措施的动物模型。
  • 图  1  PBOO术结扎方法

    Figure  1.  Ligature method in surgical progress of PBOO

    Left: Separation of bladder neck; Right: The bladder neck is ligated by a metal rod as a guide.

    图  2  泌尿系统的大体形态病理学变化

    Figure  2.  Pathological changes of the gross morphology of the urinary system

    A: Gross morphology of bladder, ureter, pelvis and kidney; single-headed arrow indicates ureter, double-headed arrow indicates bladder; B: Retrograde urography; C: Gross morphology of kidney; D: Renal parenchymal thickness (n=12), ****P<0.000 1.

    图  3  肾皮质的组织学改变

    Figure  3.  Histopathological changes of kidney cortex

    图  4  肾髓质的组织学改变

    Figure  4.  Histopathological changes of kidney medulla

    图  5  肾脏组织中Vimentin、PCNA和α-SMA的蛋白表达水平变化

    Figure  5.  Protein expression levels of Vimentin, PCNA and α-SMA in the kidney tissue of the control group, the sham operation group and the PBOO group

    n=12. ****P<0.000 1.

    图  6  免疫组化染色检测PCNA表达水平

    Figure  6.  Immunohistochemistry staining results of PCNA

    图  7  免疫荧光染色检测α-SMA表达水平。×200

    Figure  7.  Immunohistochemistry staining results of α-SMA. ×200

    表  1  对照组、假手术组与PBOO组的肾脏质量

    Table  1.   Kidney mass of the control, sham operation and PBOO groups

    Groupnmleft kidney/mgmright kidney/mg
    Control 12 92.06±5.06 89.83±4.45
    Sham operation 12 85.78±2.91 85.93±3.94
    PBOO 12 131.10±6.92*, # 137.40±9.81*, #
     * P<0.001, vs. control group; # P<0.000 1, vs. sham operation group.
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出版历程
  • 收稿日期:  2020-12-25
  • 修回日期:  2021-07-05
  • 网络出版日期:  2021-12-06
  • 刊出日期:  2021-09-20

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