超声造影对兔糖尿病肾病模型早期微循环血流灌注变化的实验研究

Experimental study of contrast-enhanced ultrasound to evaluate the changes of microcirculation blood perfusion in early stage of rabbit diabetic nephropathy model

目的 探讨分析兔糖尿病肾病模型早期微循环血流灌注变化。方法 研究对象为80只健康雄性新西兰兔,随机分为对照组(健康兔)和糖尿病肾病组。经耳缘静脉快速注射2.5%四氧嘧啶溶液(100 mg/kg)建立糖尿病兔模型,同时对照组兔子注射同剂量生理盐水。建模成功1周、2周、4周、6周、8周、10周、12周后随机处死一只兔,取双肾作病理标本。两组兔分别在造模结束后3天、1周、2周、4周、6周、8周、10周、12周均经耳缘静脉团注造影剂,后迅速注射生理盐水1.5 ml冲洗管道,进行实时超声造影,造影结束后,处死兔行Masson染色,观察病理改变。利用时间-强度定量分析软件分析两组兔不同阶段肾皮质的血流灌注变化。结果 时间-强度曲线参数:建模1周、2周后峰值强度呈上升趋势,曲线下面积(AUC)增大、达峰时间(TTP)减少,曲线增强上升斜率(AS)增大,建模4周、6周后,峰值强度、AUC均较对照组升高,TTP、AS均与对照组类似,建模8周、10周、12周,TTP增多,AS减缓,峰值强度减半时间增长、峰值强度下降。病理结果显示1周、2周肾小球增大,系膜基质、基底膜无异常;4周、6周系膜细胞上升;8周、10周、12周肾小球系膜细胞增多、基底膜厚度增加,大部分毛细血管管腔闭塞,部分肾小球硬化。结论 超声造影作为一项简单方便的检查技术,可对早期兔糖尿病肾病早期微循环血流灌注变化展开定量分析,能够有效反映肾脏病理改变,为糖尿病肾病早期治疗提供依据。

Objective To analyze the changes of microcirculation blood perfusion in the early stage of rabbit diabetic nephropathy model.Methods 80 healthy male New Zealand rabbits were randomly divided into healthy group and diabetic nephropathy group.The diabetic rabbit model was established by rapid injection of 2.5%alloxouracil solution(100 mg/kg)through the auricular vein,while the healthy rabbits were injected with the same dose of normal saline.After 1,2,4,6,8,10 and 12 weeks of successful modeling,one rabbit was randomly killed and both kidneys were taken as pathological specimens.At 3 days,1 week,2 weeks,4 weeks,6 weeks,8 weeks,10 weeks and 12 weeks after the end of modeling,rabbits in both groups were injected with contrast agent through the ear margin vein mass,and then 1.5 ml of normal saline was quickly injected into the irrigation pipeline to perform real-time CEUS.After the end of imaging,rabbits were killed and Masson staining was performed to observe the pathological changes.The changes of blood perfusion in renal cortex of two groups were analyzed by time-intensity quantitative analysis software.Results Time-intensity curve parameters:After 1 and 2 weeks of modeling,the peak intensity showed an upward trend,the area under the curve(AUC)increased,the peak time to reach the peak(TTP)decreased,and the curve enhancement slope(AS)increased.After 4 and 6 weeks of modeling,the peak intensity and AUC were both higher than that of the control group,and TTP and AS were similar to the control group.After 8,10 and 12 weeks of modeling,TTP increased.AS slowed down,the peak intensity halved,and the peak intensity decreased.Pathological results showed,at 1 week,2 weeks,glomerular enlargement,mesangial matrix,basement membrane had no abnormality;at 4 weeks,6 weeks,mesangial cells increased;at 8 weeks,10 weeks,12 weeks,mesangial cells increased,basement membrane thickness increased,most of the capillary lumen blocked,part of the glomerular sclerosis.Conclusion As a simple and convenient examination technique,contrast-enhanced ultrasound can quantitatively analyze the changes of microcirculation blood perfusion in the early stage of rabbit diabetic nephropathy,which can effectively reflect the pathological changes of kidney and provide evidence for the early treatment of diabetic nephropathy.