CLARITY技术在小鼠急性肾损伤模型中的应用

Application of CLARITY technique in a mouse model of acute kidney injury

目的基于小鼠急性肾损伤模型探讨CLARITY技术在病理诊断中应用的价值。方法将24只昆明小鼠随机均分为实验组和对照组,每组(各12只)分别建立左侧输尿管完全梗阻模型,0d、2d、4d、6d后每次取3只实验组小鼠进行透明化处理,对照组行常规HE染色。联合应用肉眼观察法、激光共聚焦显微镜观察法和常规显微镜观察法检测肾脏组织的形态学改变。采用TissueFAXS^(TM) Cytometry组织细胞定量分析系统对共聚焦显微镜二维图片中的肾小管上皮细胞数量和肾小管管腔面积进行定量分析。结果肉眼观梗阻侧肾脏,随着时间延长,肾体积增大、充血、水肿,肾盂和输尿管扩张,肾实质结构逐渐萎缩,肾盏融合,肾皮质开始变薄;实验组和对照组均观察到肾脏梗阻后肾小管上皮细胞扁平化、扩张,随着梗阻时间延长,逐渐出现肾小管萎缩消失、组织结构破坏,典型形态消失,纤维结缔组织增生,肾脏出现纤维化;实验组采用激光共聚焦显微镜扫描发现同一个组织的不同层面肾小管上皮细胞数量和肾小管管腔面积分布明显不同。结论透明化技术可观察急性肾损伤后肾脏不同组织层面的病理改变,比HE染色的二维层面提供更多的信息,在病理学和形态学研究领域有较高的应用价值。

Objective To explore the value of CLARITY in pathological diagnosis based on acute renal injury in mice models. Methods Twenty-four mice from Kunming were randomly divided into the experimental group(n=12)and the control group(n=12).After the complete obstruction of the left ureter models were established for each group,3 mice were selected from the experimental group each time on the 0,2nd,4th,and 6th day and treated by CLARITY technique,and the mice in control group were treated by HE staining.The morphological changes of the kidney tissue were observed by naked eyes,confocal laser scanning microscopy and conventional microscope.The quantity of tubular epithelial cells and lumen area of the tubules in the two-dimensional image of the confocal laser scanning microscopy were quantified by using the TissueFAXS TM Cytometry tissue quantitative analysis system. Results As time went on,the naked eye observation showed that the kidney in the obstructed side was enlarged,hyperemia and edema,the renal pelvis and ureter were dilated,the renal parenchyma was atrophied gradually,the renal calyces were fused,and the renal cortex became thinner.In the experimental group and the control group,it was found that the renal tubular epithelial cells were flattened and dilated after renal obstruction.As the obstruction time prolonged,renal tubular was atrophied and tissues disappeared,the tissue structure was destroyed,the typical morphology disappeared,the fibrotic connective tissues proliferated and the kidney appeared fibrosis.Laser confocal microscopy revealed significant differences in the number of renal tubular epithelial cells and the distribution of renal tubular lumen in different layers of the same tissue. Conclusion CLARITY technique can observe the pathological changes of different tissue depth of kidney after acute kidney injury,and it can present more data than the two-dimentional layer of HE staining,and has high application value in the field of pathology and morphology.