Wang Zhao-Ying, Zhang Zhao-Xia, Jin Li-Ming, Liu Bo, Shen Lian-Ju, He Da-Wei, Wei Guang-Hui
Key Laboratory of Children Urogenital Development and Tissue Engineering, Key Laboratory of Children's Development and Disorders of the Ministry of Education, National Clinical Research Center for Children's Health and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of Pediatrics, Children's Hospital of Chongqing Medical University, Chongqing 400014, China.
Department of Urology, Children's Hospital of Chongqing Medical University, Chongqing 400014, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2021 Sep;52(5):825-831. doi: 10.12182/20210960502.
To establish an animal model of reflux renal damage through bladder outlet obstruction.
Sixty male C57BL/6 mice aged 6-8 weeks were randomly assigned to a control group, a sham operation group, and a partial bladder outlet obstruction (PBOO) group, with 20 mice in each group. Laparotomy were performed on the PBOO mice under anesthesia in order to separate the bladder necks and to perform guided partial ligation of the bladder neck with a metal rod of 0.3 mm diameter. Mice in the sham operation group had laparotomy and had their bladder necks separated without ligation. The control group did not receive any treatment. 7 days after the surgery, 12 surviving mice were randomly selected from each group to observe the general changes of the bladder, ureter, renal pelvis and kidney. Retrograde urography was performed through the bladder. Kidney tissues were extracted for histopathological analysis. The expression levels of Vimentin, proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA) were examined with Western blot, immunohistochemistry and immunofluorescence staining tests, respectively.
Compared with the control and sham operation group, the bladder, ureter, and renal pelvis of the mice in the PBOO group were significantly enlarged, vesicoureteral reflux was more obvious, the kidney volume and mass increased ( <0.001), and renal parenchyma became thinner ( <0.000 1). Histopathological staining showed glomerular atrophy, renal tubule expansion, tubulointerstitial inflammatory cell infiltration, glomerular basement membrane hyperplasia and obvious interstitial fibrosis. Western blot, immunofluorescence and immunohistochemistry staining showed that the expression levels of Vimentin, PCNA and α-SMA in kidney tissue were elevated ( <0.000 1).
After PBOO, the bladder, ureter, and kidney of the mice showed obvious morphological alteration and presented reflux renal fibrosis-like damage. This can be used as an animal model to study the pathological alteration mechanism and therapeutic measures of renal fibrosis caused by bladder outlet obstruction.
通过膀胱出口梗阻建立反流性肾损伤动物模型。
将60只6 - 8周龄的雄性C57BL/6小鼠随机分为对照组、假手术组和部分膀胱出口梗阻（PBOO）组，每组20只。对PBOO组小鼠在麻醉下进行剖腹手术，分离膀胱颈并用直径0.3 mm的金属棒进行引导下的膀胱颈部分结扎。假手术组小鼠进行剖腹手术，分离膀胱颈但不结扎。对照组不接受任何处理。术后7天，每组随机选取12只存活小鼠，观察膀胱、输尿管、肾盂和肾脏的大体变化。通过膀胱进行逆行尿路造影。提取肾脏组织进行组织病理学分析。分别采用蛋白质免疫印迹法、免疫组织化学法和免疫荧光染色法检测波形蛋白（Vimentin）、增殖细胞核抗原（PCNA）和α - 平滑肌肌动蛋白（α - SMA）的表达水平。
与对照组和假手术组相比，PBOO组小鼠的膀胱、输尿管和肾盂明显增大，膀胱输尿管反流更明显，肾脏体积和重量增加（<0.001），肾实质变薄（<0.000 1）。组织病理学染色显示肾小球萎缩、肾小管扩张、肾小管间质炎性细胞浸润、肾小球基底膜增生和明显的间质纤维化。蛋白质免疫印迹法、免疫荧光法和免疫组织化学染色显示肾脏组织中Vimentin、PCNA和α - SMA的表达水平升高（<0.000 1）。
PBOO后，小鼠的膀胱、输尿管和肾脏出现明显的形态学改变，并呈现反流性肾纤维化样损伤。这可作为研究膀胱出口梗阻所致肾纤维化的病理改变机制及治疗措施的动物模型。
