小鼠单侧输尿管梗阻模型肾损伤随时间变化的研究  

Study about renal injury of unilateral ureteral obstruction model with time

作　　者：李禹蒙 贾西贝[1] 马静[1] 邢丽[1] Li Yumeng;Jia Xibei;Ma Jing;Xing Li(Department of Nephrology,the First Affiliated Hospital of Harbin Medical University,Harbin 150001,China)

机构地区：[1]哈尔滨医科大学第一附属医院肾内一科,哈尔滨150001

出　　处：《中国医师杂志》2022年第10期1468-1473,1478,共7页Journal of Chinese Physician

基　　金：国家自然科学基金(82000646);黑龙江省自然科学基金(LH2019H081);黑龙江省博士后科研启动金(LBH-Q19152);哈尔滨医科大学附属第一医院科研创新基金(2020M26)

摘　　要：目的建立小鼠单侧(左侧)输尿管梗阻(UUO)模型,探究肾损伤随时间的变化情况和相关机制。方法50只小鼠随机分为两组:假手术组及UUO模型组(结扎左侧输尿管制成UUO模型),观察不同时间点两组生化指标、左肾重量/终末体重(LR/BW)、右肾重量/终末体重(RR/BW),并计算左肾重量/右肾重量比值(LR/RR)。采用HE、Masson及PAS染色法检测小鼠的肾脏病理学变化。免疫荧光染色观察小鼠肾小管周围毛细血管(PTC)缺失、肾实质细胞增殖(Ki67^(+)细胞)以及巨噬细胞(CD68^(+)标记)、成纤维细胞浸润及肾脏Wnt/β-catenin表达情况。结果UUO组小鼠体重在造模第3天迅速下降[(18.2±1.1)g vs(22.4±1.2)g],此后体重缓慢增加直至第28天,在第60天体重显著下降[(17.5±0.8)g];LR/RR和LR/BW在第3天显著上升,随后逐渐下降;第60天时肾功能明显恶化[血清肌酐(0.89±0.09)mg/dl,尿素氮(41.26±5.65)mg/dl];UUO组梗阻侧肾脏光镜显示肾小管间质纤维化和肾小球硬化,随着时间的推移,PTC缺失逐渐加重;巨噬细胞最初在左肾实质内显著增多,但在28 d后开始减少;成纤维细胞数量在梗阻侧(左侧)肾脏前14 d显著增多,此后下降至正常水平;UUO组非梗阻侧肾脏的细胞数量与假手术组差异无统计学意义;UUO组梗阻侧(左侧)Wnt/β-catenin免疫荧光强度在肾脏损伤后前14 d的表达显著上调,28 d后表达下降。结论UUO肾脏纤维化的发展涉及很多方面的变化,包括PTC缺失、巨噬细胞的浸润、成纤维细胞激活以及表达,但是这些变化随着时间而减弱。Objective The unilateral(left)ureteral obstruction(UUO)model was established in mice to explore the changes of renal injury with time and the related mechanisms.Methods Fifty mice were randomly divided into two groups:sham group and UUO group(UUO model was made by unilateral ureteral ligation).The biochemical indexes,left kidney weight/final weight(LR/BW)and right kidney weight/final weight(RR/BW)of the two groups at different time points were observed,and the left kidney weight/right kidney weight ratio(LR/RR)was calculated.Hematoxylin-eosin(HE),Masson and periodic acid-Schiff(PAS)staining were used to detect the pathological changes of the kidney in mice.Immunofluorescence staining was used to observe the loss of peritubular capillaries(PTC),proliferation of renal parenchymal cells(Ki67^(+)cells),macrophages(CD68^(+)markers),infiltration of fibroblasts and expression of Wnt/β-catenin in the kidney of mice.Results The weight of mice in UUO group decreased rapidly[(18.2±1.1)g vs(22.4±1.2)g]on the third day of modeling,then slowly increased until the 28th day,and significantly decreased[(17.5±0.8)g]on the 60th day;LR/RR and LR/BW increased significantly in the third day,and then decreased gradually;Renal function of mice in UUO group deteriorated significantly on the 60th day[serum creatinine(0.89±0.09)mg/dl,urea nitrogen(41.26±5.65)mg/dl].In UUO group,renal tubulointerstitial fibrosis and glomerulosclerosis were observed under light microscope in the obstructed kidney;with the passage of time,PTC loss gradually increased;macrophages increased significantly in the left renal parenchyma at first,but began to decrease 28 days later;the number of fibroblasts increased significantly in the first 14 days of the obstructed side(left side)kidney,and then decreased to the normal level;There was no significant difference in the cell number of the non obstructive kidney between UUO group and sham group;The immunofluorescence intensity expression of Wnt/β-catenin of obstructive side(left side)in UUO group was signific

关 键 词：输尿管梗阻 疾病模型 动物 肾间质纤维化 肾小管周围毛细血管 巨噬细胞 

分 类 号：R69[医药卫生—泌尿科学]